#include #include #include #include #include "Correct.h" #include "Levenshtein_distance.h" #include "edlib.h" #include "Assembly.h" long long T_total_match=0; long long T_total_unmatch=0; long long T_total_mis=0; pthread_mutex_t debug_statistics ; void clear_Round2_alignment(Round2_alignment* h) { clear_Correct_dumy_pure(&(h->dumy)); clear_Cigar_record(&(h->cigar)); clear_Cigar_record(&(h->tmp_cigar)); h->obtained_cigar_length = 0; } void init_Round2_alignment(Round2_alignment* h) { init_Correct_dumy(&(h->dumy)); init_Cigar_record(&(h->cigar)); init_Cigar_record(&(h->tmp_cigar)); h->obtained_cigar_length = 0; } void destory_Round2_alignment(Round2_alignment* h) { destory_Correct_dumy(&(h->dumy)); destory_Cigar_record(&(h->cigar)); destory_Cigar_record(&(h->tmp_cigar)); } ///y_length > x_length unsigned int edit_distance_normal_test_banded(char* y, int y_length, char* x, int x_length, int error_cut, int matrix[1000][1000] ) { memset(matrix, 0, sizeof(matrix)); int i, j; for (i = 0; i <= x_length; i++) { matrix[i][0] = i; } int digonal, up, left; unsigned int min; ///一列列算的 for (i = 0; i < x_length; i++) { for (j = 0; j < y_length; j++) { ///matrix[i + 1][j + 1] digonal = matrix[i][j] + (x[i] != y[j]); up = matrix[i + 1][j] + 1; left = matrix[i][j + 1] + 1; min = digonal; if (up < min) { min = up; } if (left< min) { min = left; } matrix[i + 1][j + 1] = min; } } min = (unsigned int)-1; for (j = x_length; j <= y_length; j++) { if (matrix[i][j] < min) { min = matrix[i][j]; } } return min <= error_cut?min:(unsigned int)(-1); } void verify_get_interval(long long window_start, long long window_end, overlap_region_alloc* overlap_list,Correct_dumy* dumy) { long long i; long long match_length = 0; long long match_lengthNT = 0; long long Len; for (i = 0; i < overlap_list->length; i++) { if((Len = OVERLAP(window_start, window_end, overlap_list->list[i].x_pos_s, overlap_list->list[i].x_pos_e)) > 0) { if (Len == WINDOW) { match_length++; long long j; for (j = 0; j < dumy->length; j++) { if (i==dumy->overlapID[j]) { break; } } if (j >= dumy->length) { fprintf(stderr, "+ERROR interval\n"); fprintf(stderr, "i: %u, window_start: %u, window_end: %u, x_pos_s: %u, x_pos_e: %u\n", i, window_start, window_end, overlap_list->list[i].x_pos_s, overlap_list->list[i].x_pos_e); } } else { match_lengthNT++; long long j; for (j = 0; j < dumy->lengthNT; j++) { if (i==dumy->overlapID[dumy->size - j - 1]) { break; } } if (j >= dumy->lengthNT) { fprintf(stderr, "-ERROR interval\n"); fprintf(stderr, "i: %u, window_start: %u, window_end: %u, x_pos_s: %u, x_pos_e: %u, dumy->lengthNT: %u\n", i, window_start, window_end, overlap_list->list[i].x_pos_s, overlap_list->list[i].x_pos_e, dumy->lengthNT); } } } } if (match_length != dumy->length || match_lengthNT != dumy->lengthNT) { fprintf(stderr, "****************ERROR interval length*******************\n"); fprintf(stderr, "match_length: %u\n", match_length); fprintf(stderr, "dumy->length: %u\n", dumy->length); fprintf(stderr, "window_start: %u, window_end: %u\n", window_start, window_end); } } inline int get_interval_back(long long window_start, long long window_end, overlap_region_alloc* overlap_list, Correct_dumy* dumy) { long long i; int flag = 0; for (i = dumy->start_i; i < overlap_list->length; i++) { ///只会发生在这个interval比list里所有元素都小的情况 ///这种情况下一个interval需要从0开始 if (window_start < overlap_list->list[i].x_pos_s) { dumy->start_i = 0; dumy->length = 0; return -1; } else if(window_start >= overlap_list->list[i].x_pos_s && window_start <= overlap_list->list[i].x_pos_e) { dumy->start_i = i; break; } } ///只会发生在这个window比list里所有元素都大的情况 ///这种情况下一个window也无需遍历了 if (i >= overlap_list->length) { dumy->start_i = overlap_list->length; dumy->length = 0; return -2; } ///走到这里的时候,至少window_start的要求是满足了 dumy->length = 0; for (; i < overlap_list->length; i++) { if(overlap_list->list[i].x_pos_s <= window_start && overlap_list->list[i].x_pos_e >= window_end) { dumy->overlapID[dumy->length] = i; dumy->length++; } else if(overlap_list->list[i].x_pos_s > window_start) { break; } } if ( dumy->length == 0) { return 0; } else { return 1; } } inline int get_interval(long long window_start, long long window_end, overlap_region_alloc* overlap_list, Correct_dumy* dumy) { long long i; int flag = 0; long long Begin, End, Len; for (i = dumy->start_i; i < overlap_list->length; i++) { ///只会发生在这个interval比list里所有元素都小的情况 ///这种情况下一个interval需要从0开始 if (window_end < overlap_list->list[i].x_pos_s) { dumy->start_i = 0; dumy->length = 0; return 0; } else ///只要window_end >= overlap_list->list[i].x_pos_s,就有可能重叠 { dumy->start_i = i; break; } } ///只会发生在这个window比list里所有元素都大的情况 ///这种情况下一个window也无需遍历了 if (i >= overlap_list->length) { dumy->start_i = overlap_list->length; dumy->length = 0; return -2; } dumy->length = 0; dumy->lengthNT = 0; for (; i < overlap_list->length; i++) { if((Len = OVERLAP(window_start, window_end, overlap_list->list[i].x_pos_s, overlap_list->list[i].x_pos_e)) > 0) { if (Len == WINDOW) { dumy->overlapID[dumy->length] = i; dumy->length++; } else { dumy->lengthNT++; dumy->overlapID[dumy->size - dumy->lengthNT] = i; } } if(overlap_list->list[i].x_pos_s > window_end) { break; } } if ( dumy->length + dumy->lengthNT == 0) { return 0; } else { return 1; } } inline int get_available_interval(long long window_start, long long window_end, overlap_region_alloc* overlap_list, Correct_dumy* dumy) { long long i; int flag = 0; long long Begin, End, Len; long long overlap_length; for (i = dumy->start_i; i < overlap_list->length; i++) { ///只会发生在这个interval比list里所有元素都小的情况 ///这种情况下一个interval需要从0开始 if (window_end < overlap_list->list[i].x_pos_s) { dumy->start_i = 0; dumy->length = 0; dumy->lengthNT = 0; return 0; } else ///只要window_end >= overlap_list->list[i].x_pos_s,就有可能重叠 { dumy->start_i = i; break; } } ///只会发生在这个window比list里所有元素都大的情况 ///这种情况下一个window也无需遍历了 if (i >= overlap_list->length) { dumy->start_i = overlap_list->length; dumy->length = 0; dumy->lengthNT = 0; return -2; } dumy->length = 0; dumy->lengthNT = 0; long long fake_length = 0; for (; i < overlap_list->length; i++) { ///是否重叠 if((Len = OVERLAP(window_start, window_end, overlap_list->list[i].x_pos_s, overlap_list->list[i].x_pos_e)) > 0) { ///重叠数量 fake_length++; ///重叠是否有效 overlap_length = overlap_list->list[i].x_pos_e - overlap_list->list[i].x_pos_s + 1; if (overlap_length * OVERLAP_THRESHOLD <= overlap_list->list[i].align_length) { dumy->overlapID[dumy->length] = i; dumy->length++; } } if(overlap_list->list[i].x_pos_s > window_end) { break; } } ///fake_length是重叠的数量,而不是有效重叠的数量 if (fake_length == 0) { return 0; } else { return 1; } } ///Len = OVERLAP(window_start, window_end, overlap_list->list[i].x_pos_s, overlap_list->list[i].x_pos_e)) void print_string(char* s, int l) { for (size_t i = 0; i < l; i++) { fprintf(stderr, "%c", s[i]); } fprintf(stderr, "\n"); } void test_edit_distance_by_edlib(char* x_string, char* y_string, long long x_len, long long o_len, int threashold, int error, long long* total_mis) { EdlibAlignResult result = edlibAlign(x_string, x_len, y_string, o_len, edlibNewAlignConfig(threashold, EDLIB_MODE_HW, EDLIB_TASK_PATH, NULL, 0)); if (result.status == EDLIB_STATUS_OK) { if (result.editDistance != error) { (*total_mis)++; if ((int)error != -1 && result.editDistance==-1) { fprintf(stderr, "ERROR1\n"); } if ((int)error < result.editDistance==-1 && (int)error != -1 && result.editDistance!=-1) { fprintf(stderr, "ERROR2\n"); } int up_length = o_len - result.endLocations[0] - 1; int left_length = result.endLocations[0] - x_len; char* cigar = edlibAlignmentToCigar(result.alignment, result.alignmentLength, EDLIB_CIGAR_STANDARD); int cigar_length = strlen(cigar); int i = cigar_length - 1; int j = 0; char tmp; while (i >= 0) { if (up_length < 0 || left_length < 0) { break; } if (cigar[i] == 'I' || cigar[i] == 'D') { tmp = cigar[i]; cigar[i] = '\0'; j = i - 1; while (cigar[j] <= '9' && cigar[j] >= '0' && j >= 0) { j--; } j++; int Len = atoi(cigar + j); cigar[i] = tmp; i = j - 1; if (tmp == 'I') { up_length = up_length - Len; left_length = left_length + Len; } else { left_length = left_length - Len; up_length = up_length + Len; } } else { i--; } } if (up_length >= 0 && left_length >= 0) { fprintf(stderr, "****\nedlib: %d, alignmentLength: %d, startLocations: %d, endLocations: %d\n", result.editDistance, result.alignmentLength, result.startLocations[0], result.endLocations[0]); fprintf(stderr,"%s\n", cigar); print_string(x_string, x_len); print_string(y_string, o_len); fprintf(stderr, "BPM: %d\n", error); } free(cigar); } } edlibFreeAlignResult(result); } void fill_subregion(char* r, long long start_pos, long long length, uint8_t strand, All_reads* R_INF, long long ID, int extra_begin, int extra_end) { recover_UC_Read_sub_region(r+extra_begin, start_pos, length, strand, R_INF, ID); memset(r, 'N', extra_begin); memset(r+extra_begin+length, 'N', extra_end); } void determine_overlap_region(int threshold, long long y_start, long long y_ID, long long Window_Len, All_reads* R_INF, int* r_extra_begin, int* r_extra_end, long long* r_y_start, long long* r_y_length) { int extra_begin; int extra_end; long long currentIDLen; long long o_len; extra_begin = extra_end = 0; ///y maybe less than 0 y_start = y_start - threshold; ///the length of y currentIDLen = Get_READ_LENGTH((*R_INF), y_ID); o_len = MIN(Window_Len, currentIDLen - y_start); extra_end = Window_Len - o_len; if (y_start < 0) { extra_begin = -y_start; y_start = 0; o_len = o_len - extra_begin; } (*r_extra_begin) = extra_begin; (*r_extra_end) = extra_end; (*r_y_start) = y_start; (*r_y_length) = o_len; } void verify_window(long long window_start, long long window_end, overlap_region_alloc* overlap_list,Correct_dumy* dumy, All_reads* R_INF, char* r_string) { long long i; long long currentID, currentIDLen; long long x_start, y_start, o_len; long long Window_Len = WINDOW + (THRESHOLD << 1); char* x_string = NULL; char* y_string = NULL; long long x_end, x_len; int end_site; unsigned int error; int groupLen = 0; int return_sites[GROUP_SIZE]; unsigned int return_sites_error[GROUP_SIZE]; uint64_t overlapID[GROUP_SIZE]; uint64_t y_startGroup[GROUP_SIZE]; int y_extra_begin[GROUP_SIZE]; int y_extra_end[GROUP_SIZE]; int extra_begin; int extra_end; ///这些是整个window被完全覆盖的 for (i = 0; i < dumy->length; i++) { extra_begin = extra_end = 0; ///整个window被覆盖的话,read本身上的区间就是[window_start, window_end] x_len = WINDOW; currentID = dumy->overlapID[i]; x_start = window_start; ///y上的相对位置 y_start = (x_start - overlap_list->list[currentID].x_pos_s) + overlap_list->list[currentID].y_pos_s; determine_overlap_region(THRESHOLD, y_start, overlap_list->list[currentID].y_id, Window_Len, R_INF, &extra_begin, &extra_end, &y_start, &o_len); fill_subregion(dumy->overlap_region_group[groupLen], y_start, o_len, overlap_list->list[currentID].y_pos_strand, R_INF, overlap_list->list[currentID].y_id, extra_begin, extra_end); y_extra_begin[groupLen] = extra_begin; y_extra_end[groupLen] = extra_end; overlapID[groupLen] = currentID; y_startGroup[groupLen] = y_start; x_string = r_string + x_start; groupLen++; if (groupLen == GROUP_SIZE) { Reserve_Banded_BPM_4_SSE_only(dumy->overlap_region_group[0], dumy->overlap_region_group[1], dumy->overlap_region_group[2], dumy->overlap_region_group[3], Window_Len, x_string, WINDOW, return_sites, return_sites_error, THRESHOLD, dumy->Peq_SSE); groupLen = 0; if (return_sites_error[0]!=(unsigned int)-1) { overlap_list->list[overlapID[0]].align_length += x_len; append_window_list(&overlap_list->list[overlapID[0]], window_start, window_end, y_startGroup[0], y_startGroup[0] + return_sites[0], (int)return_sites_error[0], y_extra_begin[0], y_extra_end[0]); } else { append_window_list(&overlap_list->list[overlapID[0]], window_start, window_end, y_startGroup[0], -1, -1, y_extra_begin[0], y_extra_end[0]); } if (return_sites_error[1]!=(unsigned int)-1) { overlap_list->list[overlapID[1]].align_length += x_len; append_window_list(&overlap_list->list[overlapID[1]], window_start, window_end, y_startGroup[1], y_startGroup[1] + return_sites[1], (int)return_sites_error[1], y_extra_begin[1], y_extra_end[1]); } else { append_window_list(&overlap_list->list[overlapID[1]], window_start, window_end, y_startGroup[1], -1, -1, y_extra_begin[1], y_extra_end[1]); } if (return_sites_error[2]!=(unsigned int)-1) { overlap_list->list[overlapID[2]].align_length += x_len; append_window_list(&overlap_list->list[overlapID[2]], window_start, window_end, y_startGroup[2], y_startGroup[2] + return_sites[2], (int)return_sites_error[2], y_extra_begin[2], y_extra_end[2]); } else { append_window_list(&overlap_list->list[overlapID[2]], window_start, window_end, y_startGroup[2], -1, -1, y_extra_begin[2], y_extra_end[2]); } if (return_sites_error[3]!=(unsigned int)-1) { overlap_list->list[overlapID[3]].align_length += x_len; append_window_list(&overlap_list->list[overlapID[3]], window_start, window_end, y_startGroup[3], y_startGroup[3] + return_sites[3], (int)return_sites_error[3], y_extra_begin[3], y_extra_end[3]); } else { append_window_list(&overlap_list->list[overlapID[3]], window_start, window_end, y_startGroup[3], -1, -1, y_extra_begin[3], y_extra_end[3]); } } } if (groupLen == 1) { end_site = Reserve_Banded_BPM(dumy->overlap_region_group[0], Window_Len, x_string, WINDOW, THRESHOLD, &error); if (error!=(unsigned int)-1) { overlap_list->list[overlapID[0]].align_length += x_len; append_window_list(&overlap_list->list[overlapID[0]], window_start, window_end, y_startGroup[0], y_startGroup[0] + end_site, (int)error, y_extra_begin[0], y_extra_end[0]); } else { append_window_list(&overlap_list->list[overlapID[0]], window_start, window_end, y_startGroup[0], -1, -1, y_extra_begin[0], y_extra_end[0]); } } else if (groupLen > 1) { Reserve_Banded_BPM_4_SSE_only(dumy->overlap_region_group[0], dumy->overlap_region_group[1], dumy->overlap_region_group[2], dumy->overlap_region_group[3], Window_Len, x_string, WINDOW, return_sites, return_sites_error, THRESHOLD, dumy->Peq_SSE); for (i = 0; i < groupLen; i++) { if (return_sites_error[i]!=(unsigned int)-1) { overlap_list->list[overlapID[i]].align_length += x_len; append_window_list(&overlap_list->list[overlapID[i]], window_start, window_end, y_startGroup[i], y_startGroup[i] + return_sites[i], (int)return_sites_error[i], y_extra_begin[i], y_extra_end[i]); } else { append_window_list(&overlap_list->list[overlapID[i]], window_start, window_end, y_startGroup[i], -1, -1, y_extra_begin[i], y_extra_end[i]); } } groupLen = 0; } long long reverse_i = dumy->size - 1; int threshold; ///这些是整个window被部分覆盖的 for (i = 0; i < dumy->lengthNT; i++) { extra_begin = extra_end = 0; currentID = dumy->overlapID[reverse_i--]; x_start = MAX(window_start, overlap_list->list[currentID].x_pos_s); x_end = MIN(window_end, overlap_list->list[currentID].x_pos_e); ///这个是和当前窗口重叠的长度 x_len = x_end - x_start + 1; threshold = x_len * THRESHOLD_RATE; ///y上的相对位置 y_start = (x_start - overlap_list->list[currentID].x_pos_s) + overlap_list->list[currentID].y_pos_s; Window_Len = x_len + (threshold << 1); determine_overlap_region(threshold, y_start, overlap_list->list[currentID].y_id, Window_Len, R_INF, &extra_begin, &extra_end, &y_start, &o_len); fill_subregion(dumy->overlap_region, y_start, o_len, overlap_list->list[currentID].y_pos_strand, R_INF, overlap_list->list[currentID].y_id, extra_begin, extra_end); x_string = r_string + x_start; y_string = dumy->overlap_region; end_site = Reserve_Banded_BPM(y_string, Window_Len, x_string, x_len, threshold, &error); if (error!=(unsigned int)-1) { overlap_list->list[currentID].align_length += x_len; append_window_list(&overlap_list->list[currentID], x_start, x_end, y_start, y_start + end_site, (int)error, extra_begin, extra_end); } else { append_window_list(&overlap_list->list[currentID], x_start, x_end, y_start, -1, -1, extra_begin, extra_end); } } } void debug_stats(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, Correct_dumy* dumy, UC_Read* overlap_read, long long* matched_overlap_0, long long* matched_overlap_1) { long long j; long long Len_x; int threshold; long long y_start; long long Len_y; long long currentIDLen; for (j = 0; j < overlap_list->length; j++) { Len_x = overlap_list->list[j].x_pos_e - overlap_list->list[j].x_pos_s + 1; if (Len_x * OVERLAP_THRESHOLD <= overlap_list->list[j].align_length) { if (overlap_list->list[j].y_pos_strand == 0) { (*matched_overlap_0)++; ///(*matched_overlap_0) = (*matched_overlap_0) + overlap_list->list[j].align_length; ///(*matched_overlap_0) = (*matched_overlap_0) + Len_x; } else { (*matched_overlap_1)++; ///(*matched_overlap_1) = (*matched_overlap_1) + overlap_list->list[j].align_length; ///(*matched_overlap_1) = (*matched_overlap_1) + Len_x; } /** threshold = Len_x * 0.04; y_start = overlap_list->list[j].y_pos_s - threshold; if (y_start < 0) { y_start = 0; } ///当前y的长度 currentIDLen = Get_READ_LENGTH((*R_INF), overlap_list->list[j].y_id); ///不能超过y的剩余长度 Len_y = MIN(Len_x + 2 * threshold, currentIDLen - y_start); if (overlap_list->list[j].y_pos_strand == 0) { recover_UC_Read(overlap_read, R_INF, overlap_list->list[j].y_id); (*matched_overlap_0)++; } else { recover_UC_Read_RC(overlap_read, R_INF, overlap_list->list[j].y_id); (*matched_overlap_1)++; } EdlibAlignResult result = edlibAlign(g_read->seq + overlap_list->list[j].x_pos_s, Len_x, overlap_read->seq + y_start, Len_y, edlibNewAlignConfig(threshold, EDLIB_MODE_HW, EDLIB_TASK_PATH, NULL, 0)); if (result.status == EDLIB_STATUS_OK && result.editDistance != -1) { char* cigar = edlibAlignmentToCigar(result.alignment, result.alignmentLength, EDLIB_CIGAR_STANDARD); int cigar_length = strlen(cigar); free(cigar); if (overlap_list->list[j].y_pos_strand == 0) { (*potiental_matched_overlap_0)++; } else { (*potiental_matched_overlap_1)++; } // if (overlap_list->list[j].shared_seed == 1 && Len_x >= 1000) // { // (*matched_overlap_0)++; // } // if (overlap_list->list[j].shared_seed == 2 && Len_x >= 1000) // { // (*matched_overlap_1)++; // } } edlibFreeAlignResult(result); **/ } } } void output_stats(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, Correct_dumy* dumy, UC_Read* overlap_read) { long long j; long long Len_x; int threshold; long long y_start; long long Len_y; long long currentIDLen; long long whole; for (j = 0; j < overlap_list->length; j++) { Len_x = overlap_list->list[j].x_pos_e - overlap_list->list[j].x_pos_s + 1; if (Len_x * OVERLAP_THRESHOLD <= overlap_list->list[j].align_length) { ; } } } inline void generate_cigar( char* path, int path_length, window_list* result, int* start, int* end, int error) { if (error == 0) { result->cigar.C_L[0] = result->x_end - result->x_start + 1; result->cigar.C_C[0] = 0; result->cigar.length = 1; return; } int i = 0; result->cigar.length = 0; ///0 is match, 1 is mismatch, 2 is up, 3 is left char pre_ciga = 5; int pre_ciga_length = 0; for (i = 0; i < path_length; i++) { if(path[i] == 1) { path[i] = 3; (*end)--; } else { break; } } for (i = path_length - 1; i >= 0; i--) { if(path[i] == 1) { path[i] = 3; (*start)++; } else { break; } } for (i = path_length - 1; i >= 0; i--) { if (pre_ciga != path[i]) { if (pre_ciga_length != 0) { result->cigar.C_L[result->cigar.length] = pre_ciga_length; result->cigar.C_C[result->cigar.length] = pre_ciga; result->cigar.length++; } pre_ciga = path[i]; pre_ciga_length = 1; } else { pre_ciga_length++; } } if (pre_ciga_length != 0) { result->cigar.C_L[result->cigar.length] = pre_ciga_length; result->cigar.C_C[result->cigar.length] = pre_ciga; result->cigar.length++; } } int verify_cigar(char* x, int x_len, char* y, int y_len, CIGAR* cigar, int error) { int x_i, y_i, cigar_i; x_i = 0; y_i = 0; cigar_i = 0; int operation; int operationLen; int i; int cigar_error = 0; int flag_error = 0; ///0 is match, 1 is mismatch, 2 is up, 3 is left ///2是x缺字符(y多字符),而3是y缺字符(x多字符) ///while (x_i < x_len && y_i < y_len && cigar_i < cigar->length) while (cigar_i < cigar->length) { operation = cigar->C_C[cigar_i]; operationLen = cigar->C_L[cigar_i]; if (operation == 0) { for (i = 0; i < operationLen; i++) { if (x[x_i]!=y[y_i]) { fprintf(stderr, "error match\n"); flag_error = 1; } x_i++; y_i++; } } else if (operation == 1) { cigar_error += operationLen; for (i = 0; i < operationLen; i++) { if (x[x_i]==y[y_i]) { fprintf(stderr, "error mismatch, cigar_i: %d, x_i: %d, y_i: %d\n",cigar_i, x_i, y_i); flag_error = 1; } x_i++; y_i++; } }///2是x缺字符(y多字符) else if (operation == 2) { cigar_error += operationLen; y_i += operationLen; }///3是y缺字符(x多字符) else if (operation == 3) { cigar_error += operationLen; x_i += operationLen; } cigar_i++; } endloop: ///return; if (cigar_error != error) { fprintf(stderr, "error cigar_error: cigar_error: %d, error: %d\n", cigar_error, error); for (i = 0; i < cigar->length; i++) { fprintf(stderr, "%u: %u\n", cigar->C_L[i], cigar->C_C[i]); } } if (flag_error == 1) { print_string(x, x_len); print_string(y, y_len); fprintf(stderr, "x_len: %d, y_len: %d, cigar_len: %d, error: %d\n", x_len, y_len, cigar->length, error); for (i = 0; i < cigar->length; i++) { fprintf(stderr, "%u: %u\n", cigar->C_L[i], cigar->C_C[i]); } } return flag_error; } int verify_cigar_2(char* x, int x_len, char* y, int y_len, Cigar_record* cigar, int error) { int x_i, y_i, cigar_i; x_i = 0; y_i = 0; cigar_i = 0; int operation; int operationLen; int i; int cigar_error = 0; int flag_error = 0; ///0 is match, 1 is mismatch, 2 is up, 3 is left ///2是x缺字符(y多字符),而3是y缺字符(x多字符) ///while (x_i < x_len && y_i < y_len && cigar_i < cigar->length) while (cigar_i < cigar->length) { operation = Get_Cigar_Type(cigar->record[cigar_i]); operationLen = Get_Cigar_Length(cigar->record[cigar_i]); if (operation == 0) { for (i = 0; i < operationLen; i++) { if (x[x_i]!=y[y_i]) { fprintf(stderr, "error match\n"); flag_error = 1; } x_i++; y_i++; } } else if (operation == 1) { cigar_error += operationLen; for (i = 0; i < operationLen; i++) { if (x[x_i]==y[y_i]) { fprintf(stderr, "error mismatch, cigar_i: %d, x_i: %d, y_i: %d\n",cigar_i, x_i, y_i); flag_error = 1; } x_i++; y_i++; } }///2是x缺字符(y多字符) else if (operation == 2) { cigar_error += operationLen; y_i += operationLen; }///3是y缺字符(x多字符) else if (operation == 3) { cigar_error += operationLen; x_i += operationLen; } cigar_i++; } endloop: ///return; /** if (cigar_error != error) { fprintf(stderr, "error cigar_error: cigar_error: %d, error: %d\n", cigar_error, error); for (i = 0; i < cigar->length; i++) { operation = Get_Cigar_Type(cigar->record[i]); operationLen = Get_Cigar_Length(cigar->record[i]); fprintf(stderr, "%u: %u\n", operationLen, operation); } } **/ if (flag_error == 1) { print_string(x, x_len); print_string(y, y_len); fprintf(stderr, "x_len: %d, y_len: %d, cigar_len: %d, error: %d\n", x_len, y_len, cigar->length, error); for (i = 0; i < cigar->length; i++) { operation = Get_Cigar_Type(cigar->record[i]); operationLen = Get_Cigar_Length(cigar->record[i]); fprintf(stderr, "%u: %u\n", operationLen, operation); } } return flag_error; } /** ///记住等于0也要重算,虽然意义不那么大就是了 #define NEED_START_POS(x) (x.error<=0) #define GET_ERROR(x) ((x.error>=0)? x.error : x.error*-1) **/ inline void recalcate_window(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, Correct_dumy* dumy, UC_Read* overlap_read) { long long j, k, i; long long Len_x; int threshold; long long y_len; long long currentIDLen; long long matches; long long y_id; int y_strand; long long y_readLen; long long x_start; long long x_end; long long x_len; long long total_y_start; long long total_y_end; long long y_start; long long y_end; long long Window_Len; char* x_string; char* y_string; int end_site; unsigned int error; int real_y_start; long long overlap_length; int extra_begin, extra_end; long long o_len; ///j负责遍历整个overlap list for (j = 0; j < overlap_list->length; j++) { y_id = overlap_list->list[j].y_id; y_strand = overlap_list->list[j].y_pos_strand; y_readLen = Get_READ_LENGTH((*R_INF), y_id); //i负责每个overlap里面的window //倒着找 //倒着用结束位置矫正 for (i = overlap_list->list[j].w_list_length - 1; i >= 0; i--) { ///找到第一个匹配的window if(overlap_list->list[j].w_list[i].y_end != -1) { ///note!!! need notification ///total_y_start = overlap_list->list[j].w_list[i].y_end + 1; ///this is the actual end postion in ystring total_y_start = overlap_list->list[j].w_list[i].y_end - overlap_list->list[j].w_list[i].extra_begin + 1; ///k遍历匹配window右侧所有不匹配的window ///如果i匹配,则k从i+1开始 ///知道第一个匹配的window结束 for (k = i + 1; k < overlap_list->list[j].w_list_length && overlap_list->list[j].w_list[k].y_end == -1; k++) { extra_begin = extra_end = 0; ///y_start有可能大于y_readLen ///这多发于最后一个window长度仅为几,而前面一个window的结束位置也超过了y_readLen-1 ///这个时候做动态规划会给超过的部分补N if (total_y_start >= y_readLen) { break; } ///there is no problem for x x_start = overlap_list->list[j].w_list[k].x_start; x_end = overlap_list->list[j].w_list[k].x_end; x_len = x_end - x_start + 1; threshold = x_len * THRESHOLD_RATE; y_start = total_y_start; Window_Len = x_len + (threshold << 1); determine_overlap_region(threshold, y_start, y_id, Window_Len, R_INF, &extra_begin, &extra_end, &y_start, &o_len); if(o_len + threshold < x_len) { break; } fill_subregion(dumy->overlap_region, y_start, o_len, y_strand, R_INF, y_id, extra_begin, extra_end); x_string = g_read->seq + x_start; y_string = dumy->overlap_region; ///note!!! need notification end_site = Reserve_Banded_BPM(y_string, Window_Len, x_string, x_len, threshold, &error); ///error等于-1说明没匹配 if (error!=(unsigned int)-1) { overlap_list->list[j].w_list[k].cigar.length = -1; overlap_list->list[j].w_list[k].y_start = y_start; overlap_list->list[j].w_list[k].y_end = y_start + end_site; overlap_list->list[j].w_list[k].error = (int)error; ///note!!! need notification overlap_list->list[j].w_list[k].extra_begin = extra_begin; overlap_list->list[j].w_list[k].extra_end = extra_end; overlap_list->list[j].align_length += x_len; } else { break; } ///note!!! need notification ///total_y_start = y_start + end_site + 1; total_y_start = y_start + end_site - extra_begin + 1; } } } ///continue; ///i负责每个overlap里面的window ///正着找 ///用起始位置矫正 for (i = 0; i < overlap_list->list[j].w_list_length; i++) { ///找到第一个匹配的window ///首先这个window要匹配 ///其次不要是第一个window,这没意义 ///最后他之前的那个window必须是不匹配,如果之前那个window匹配,也没意义 if(overlap_list->list[j].w_list[i].y_end != -1 && i != 0 && overlap_list->list[j].w_list[i - 1].y_end == -1) { ///判断这个匹配的window的起始位置有没有被计算出来 ///如果没有,就需要重新计算 if(overlap_list->list[j].w_list[i].cigar.length == -1) { ///there is no problem for x x_start = overlap_list->list[j].w_list[i].x_start; x_end = overlap_list->list[j].w_list[i].x_end; x_len = x_end - x_start + 1; threshold = x_len * THRESHOLD_RATE; Window_Len = x_len + (threshold << 1); ///y_start is the real y_start y_start = overlap_list->list[j].w_list[i].y_start; extra_begin = overlap_list->list[j].w_list[i].extra_begin; extra_end = overlap_list->list[j].w_list[i].extra_end; o_len = Window_Len - extra_end - extra_begin; fill_subregion(dumy->overlap_region, y_start, o_len, y_strand, R_INF, y_id, extra_begin, extra_end); x_string = g_read->seq + x_start; y_string = dumy->overlap_region; ///note!!! need notification end_site = Reserve_Banded_BPM_PATH(y_string, Window_Len, x_string, x_len, threshold, &error, &real_y_start, &(dumy->path_length), dumy->matrix_bit, dumy->path, overlap_list->list[j].w_list[i].error, overlap_list->list[j].w_list[i].y_end - y_start); ///到这里y_start已经被正确计算出来了 if (error != (unsigned int)-1) { generate_cigar(dumy->path, dumy->path_length, &(overlap_list->list[j].w_list[i]), &real_y_start, &end_site, error); if(real_y_start < extra_begin || end_site >= Window_Len - extra_end) { fprintf(stderr, "\nreal_y_start: %d, extra_begin: %d\n", real_y_start, extra_begin); fprintf(stderr, "end_site: %d, Window_Len: %d, extra_end: %d\n", end_site, Window_Len, extra_end); } ///note!!! need notification ///real_y_start = y_start + real_y_start; real_y_start = y_start + real_y_start - extra_begin; overlap_list->list[j].w_list[i].y_start = real_y_start; overlap_list->list[j].w_list[i].y_end = y_start + end_site; } else { fprintf(stderr, "error\n"); } } else { real_y_start = overlap_list->list[j].w_list[i].y_start; } ///再次矫正的基础位置就是real_y_start total_y_end = real_y_start - 1; ///k遍历匹配window左侧所有不匹配的window ///如果i匹配,则k从i-1开始 ///直到第一个匹配的window结束 ///因为i!=0,所以k的大小不用担心 for (k = i - 1; k >= 0 && overlap_list->list[j].w_list[k].y_end == -1; k--) { ///there is no problem in x x_start = overlap_list->list[j].w_list[k].x_start; x_end = overlap_list->list[j].w_list[k].x_end; x_len = x_end - x_start + 1; threshold = x_len * THRESHOLD_RATE; Window_Len = x_len + (threshold << 1); if(total_y_end <= 0) { break; } ///y_start may less than 0 y_start = total_y_end - x_len + 1; determine_overlap_region(threshold, y_start, y_id, Window_Len, R_INF, &extra_begin, &extra_end, &y_start, &o_len); if(o_len + threshold < x_len) { break; } fill_subregion(dumy->overlap_region, y_start, o_len, y_strand, R_INF, y_id, extra_begin, extra_end); x_string = g_read->seq + x_start; y_string = dumy->overlap_region; ///note!!! need notification end_site = Reserve_Banded_BPM_PATH(y_string, Window_Len, x_string, x_len, threshold, &error, &real_y_start, &(dumy->path_length), dumy->matrix_bit, dumy->path, -1, -1); ///error等于-1说明没匹配 if (error!=(unsigned int)-1) { generate_cigar(dumy->path, dumy->path_length, &(overlap_list->list[j].w_list[k]), &real_y_start, &end_site, error); if(real_y_start < extra_begin || end_site >= Window_Len - extra_end) { fprintf(stderr, "\nreal_y_start: %d, extra_begin: %d\n", real_y_start, extra_begin); fprintf(stderr, "end_site: %d, Window_Len: %d, extra_end: %d\n", end_site, Window_Len, extra_end); } ///y_start has no shift, but y_end has shift overlap_list->list[j].w_list[k].y_start = y_start + real_y_start - extra_begin; overlap_list->list[j].w_list[k].y_end = y_start + end_site; overlap_list->list[j].w_list[k].error = error; overlap_list->list[j].align_length += x_len; ///note!!! need notification overlap_list->list[j].w_list[k].extra_begin = extra_begin; overlap_list->list[j].w_list[k].extra_end = extra_end; } else { break; } total_y_end = y_start + real_y_start - 1 - extra_begin; } } } } ///j负责遍历整个overlap list for (j = 0; j < overlap_list->length; j++) { y_id = overlap_list->list[j].y_id; y_strand = overlap_list->list[j].y_pos_strand; y_readLen = Get_READ_LENGTH((*R_INF), y_id); overlap_length = overlap_list->list[j].x_pos_e - overlap_list->list[j].x_pos_s + 1; ///only calculate cigar for high quality overlaps if (overlap_length * OVERLAP_THRESHOLD <= overlap_list->list[j].align_length) { for (i = 0; i < overlap_list->list[j].w_list_length; i++) { ///判断cigar是否被计算 ///没被计算过就重算 ///第一个条件是判断这个窗口是否匹配 if(overlap_list->list[j].w_list[i].y_end != -1) { if(overlap_list->list[j].w_list[i].cigar.length == -1) { ///there is no problem for x x_start = overlap_list->list[j].w_list[i].x_start; x_end = overlap_list->list[j].w_list[i].x_end; x_len = x_end - x_start + 1; threshold = x_len * THRESHOLD_RATE; Window_Len = x_len + (threshold << 1); ///y_start is the real y_start y_start = overlap_list->list[j].w_list[i].y_start; extra_begin = overlap_list->list[j].w_list[i].extra_begin; extra_end = overlap_list->list[j].w_list[i].extra_end; o_len = Window_Len - extra_end - extra_begin; fill_subregion(dumy->overlap_region, y_start, o_len, y_strand, R_INF, y_id, extra_begin, extra_end); x_string = g_read->seq + x_start; y_string = dumy->overlap_region; ///note!!! need notification end_site = Reserve_Banded_BPM_PATH(y_string, Window_Len, x_string, x_len, threshold, &error, &real_y_start, &(dumy->path_length), dumy->matrix_bit, dumy->path, overlap_list->list[j].w_list[i].error, overlap_list->list[j].w_list[i].y_end - y_start); ///到这里y_start已经被正确计算出来了 if (error != (unsigned int)-1) { generate_cigar(dumy->path, dumy->path_length, &(overlap_list->list[j].w_list[i]), &real_y_start, &end_site, error); if(real_y_start < extra_begin || end_site >= Window_Len - extra_end) { fprintf(stderr, "\nreal_y_start: %d, extra_begin: %d\n", real_y_start, extra_begin); fprintf(stderr, "end_site: %d, Window_Len: %d, extra_end: %d\n", end_site, Window_Len, extra_end); } ///note!!! need notification ///real_y_start = y_start + real_y_start; real_y_start = y_start + real_y_start - extra_begin; overlap_list->list[j].w_list[i].y_start = real_y_start; overlap_list->list[j].w_list[i].y_end = y_start + end_site - extra_begin; } else { fprintf(stderr, "error\n"); } } else { overlap_list->list[j].w_list[i].y_end -= overlap_list->list[j].w_list[i].extra_begin; } } } } } /** ///j负责遍历整个overlap list for (j = 0; j < overlap_list->length; j++) { y_id = overlap_list->list[j].y_id; y_strand = overlap_list->list[j].y_pos_strand; y_readLen = Get_READ_LENGTH((*R_INF), y_id); overlap_length = overlap_list->list[j].x_pos_e - overlap_list->list[j].x_pos_s + 1; ///only calculate cigar for high quality overlaps if (overlap_length * OVERLAP_THRESHOLD <= overlap_list->list[j].align_length) { for (i = 0; i < overlap_list->list[j].w_list_length; i++) { if(overlap_list->list[j].w_list[i].y_end != -1) { ///there is no problem for x x_start = overlap_list->list[j].w_list[i].x_start; x_end = overlap_list->list[j].w_list[i].x_end; x_len = x_end - x_start + 1; x_string = g_read->seq + x_start; y_start = overlap_list->list[j].w_list[i].y_start; y_end = overlap_list->list[j].w_list[i].y_end; y_len = y_end - y_start + 1; recover_UC_Read_sub_region(dumy->overlap_region, y_start, y_len, y_strand, R_INF, y_id); y_string = dumy->overlap_region; if(verify_cigar(x_string, x_len, y_string, y_len, &overlap_list->list[j].w_list[i].cigar, overlap_list->list[j].w_list[i].error)) { fprintf(stderr, "j: %d, i: %d, y_id: %d, y_start: %d, y_end: %d\n", j, i, y_id, y_start, y_end); } } } } } **/ } inline void add_base_to_correct_read_directly(Correct_dumy* dumy, char base) { if (dumy->corrected_read_length + 2 > dumy->corrected_read_size) { dumy->corrected_read_size = dumy->corrected_read_size * 2; dumy->corrected_read = (char*)realloc(dumy->corrected_read, dumy->corrected_read_size); } dumy->corrected_read[dumy->corrected_read_length] = base; dumy->corrected_read_length++; dumy->corrected_read[dumy->corrected_read_length] = '\0'; } inline void add_base_to_correct_read(Correct_dumy* dumy, char base, int is_error) { ///deletion就不要管 if (base != 'D') { if (dumy->corrected_read_length + 2 > dumy->corrected_read_size) { dumy->corrected_read_size = dumy->corrected_read_size * 2; dumy->corrected_read = (char*)realloc(dumy->corrected_read, dumy->corrected_read_size); } dumy->corrected_read[dumy->corrected_read_length] = base; dumy->corrected_read_length++; dumy->corrected_read[dumy->corrected_read_length] = '\0'; } if (is_error) { dumy->corrected_base++; } } inline void add_segment_to_correct_read(Correct_dumy* dumy, char* segment, long long segment_length) { if (dumy->corrected_read_length + segment_length + 2 > dumy->corrected_read_size) { dumy->corrected_read_size = dumy->corrected_read_length + segment_length + 2; dumy->corrected_read = (char*)realloc(dumy->corrected_read, dumy->corrected_read_size); } memcpy(dumy->corrected_read + dumy->corrected_read_length, segment, segment_length); dumy->corrected_read_length += segment_length; dumy->corrected_read[dumy->corrected_read_length] = '\0'; } ///返回下一个backbone节点上的ID long long inline add_path_to_correct_read(Graph* backbone, Correct_dumy* dumy, long long currentNodeID, long long type, long long edgeID, Cigar_record* current_cigar, char* self_string) { //long long i; long long nodeID; ///Note: currentNodeID must be a backbone node ///currentNodeID = 0 means a fake node ///currentNodeID = i means self_string[i - 1] ///包括匹配和误配两种情况 if (type == MISMATCH) { ///这是match的情况 if(backbone->g_nodes.list[currentNodeID].mismatch_edges.list[edgeID].length == 0) { nodeID = backbone->g_nodes.list[currentNodeID].mismatch_edges.list[edgeID].out_node; add_base_to_correct_read_directly(dumy, backbone->g_nodes.list[nodeID].base); ///match所以dumy->corrected_base不要+1 ///nodeID = i means self_string[i - 1] ///add_cigar_record(self_string+nodeID-1, 1, current_cigar, 0); add_cigar_record(&(backbone->g_nodes.list[nodeID].base), 1, current_cigar, 0); /***********需要注释掉********* */ if (nodeID != currentNodeID + 1) { fprintf(stderr, "error match\n"); } /***********需要注释掉********* */ return nodeID; } else ///这是mismatch的情况 { nodeID = backbone->g_nodes.list[currentNodeID].mismatch_edges.list[edgeID].out_node; add_base_to_correct_read_directly(dumy, backbone->g_nodes.list[nodeID].base); dumy->corrected_base++; char merge_base = 0; merge_base = seq_nt6_table[(uint8_t)backbone->g_nodes.list[nodeID].base]; merge_base = merge_base << 3; ///这种中间节点只有一个元素,所以直接list[0] nodeID = backbone->g_nodes.list[nodeID].mismatch_edges.list[0].out_node; merge_base = merge_base | seq_nt6_table[(uint8_t)backbone->g_nodes.list[nodeID].base]; add_cigar_record(&merge_base, 1, current_cigar, 1); /** add_cigar_record(&(backbone->g_nodes.list[nodeID].base), 1, current_cigar, 1); nodeID = backbone->g_nodes.list[nodeID].mismatch_edges.list[0].out_node; **/ /***********需要注释掉********* */ if (nodeID != currentNodeID + 1) { fprintf(stderr, "error mismatch\n"); } /***********需要注释掉********* */ return nodeID; } } else if (type == DELETION) { nodeID = backbone->g_nodes.list[currentNodeID].deletion_edges.list[edgeID].out_node; dumy->corrected_base += nodeID - currentNodeID; ///currentNodeID = i means self_string[i - 1] add_cigar_record(self_string + currentNodeID, nodeID - currentNodeID, current_cigar, DELETION); /***********需要注释掉********* */ if (!(nodeID >= backbone->s_start_nodeID && nodeID <= backbone->s_end_nodeID)) { fprintf(stderr, "error deletion 1\n"); } if (nodeID <= currentNodeID) { fprintf(stderr, "error deletion 2\n"); } /***********需要注释掉********* */ return nodeID; } else if (type == INSERTION) { ///这个一定要变成0 backbone->g_nodes.list[currentNodeID].num_insertions = 0; nodeID = backbone->g_nodes.list[currentNodeID].insertion_edges.list[edgeID].out_node; long long step = backbone->g_nodes.list[currentNodeID].insertion_edges.list[edgeID].length; long long i; for (i = 0; i < step; i++) { add_base_to_correct_read_directly(dumy, backbone->g_nodes.list[nodeID].base); add_cigar_record(&backbone->g_nodes.list[nodeID].base, 1, current_cigar, INSERTION); ///只有一条边 nodeID = backbone->g_nodes.list[nodeID].insertion_edges.list[0].out_node; } dumy->corrected_base += step; ///currentNodeID = i means self_string[i - 1] ///add_cigar_record(self_string + currentNodeID, step, current_cigar, INSERTION); /***********需要注释掉********* */ if (nodeID != currentNodeID) { fprintf(stderr, "error insertion\n"); } /***********需要注释掉********* */ return nodeID; } else { fprintf(stderr, "error type\n"); } } void get_seq_from_Graph(Graph* backbone, Correct_dumy* dumy, Cigar_record* current_cigar, char* self_string) { long long new_seq_length = 0; long long currentNodeID; long long i; // 总共有以下几种情况: // 1. match 2. mismatch (A, C, G, T, N) 3. deletion 4. insertion (A, C, G, T) // 其实就是 1. 自己本身的weight 2. alignToNode的weight 3. insertion节点的weight long long max_count; int max_type; long long max_edge; long long total_count; long long nodeID; char current_base; long long current_weight; currentNodeID = backbone->s_start_nodeID; while (currentNodeID != backbone->s_end_nodeID) { total_count = 0; max_count = -1; max_type = -1; max_edge = -1; ///假如这是个backbone节点 ///有三类出边 ///1. mismatch_edges 2. insertion_edges 3. deletion_edges if (currentNodeID >= backbone->s_start_nodeID && currentNodeID <= backbone->s_end_nodeID) { ///mismatch_edges for (i = 0; i < backbone->g_nodes.list[currentNodeID].mismatch_edges.length; i++) { if (backbone->g_nodes.list[currentNodeID].num_insertions != 0) { current_weight = backbone->g_nodes.list[currentNodeID].mismatch_edges.list[i].weight - backbone->g_nodes.list[currentNodeID].mismatch_edges.list[i].num_insertions; } else { current_weight = backbone->g_nodes.list[currentNodeID].mismatch_edges.list[i].weight; } total_count = total_count + current_weight; ///match ///match要处理插入的情况 ///如果这里有insertion, 这个节点会过两遍 ///第一遍num_insertions > 0, 第二遍num_insertions=0 if (current_weight > max_count) { max_count = current_weight; max_edge = i; max_type = MISMATCH; } } ///insertion_edges if (backbone->g_nodes.list[currentNodeID].num_insertions != 0) { for (i = 0; i < backbone->g_nodes.list[currentNodeID].insertion_edges.length; i++) { total_count = total_count + backbone->g_nodes.list[currentNodeID].insertion_edges.list[i].weight; if (backbone->g_nodes.list[currentNodeID].insertion_edges.list[i].weight > max_count) { max_count = backbone->g_nodes.list[currentNodeID].insertion_edges.list[i].weight; max_edge = i; max_type = INSERTION; } } } ///deletion_edges for (i = 0; i < backbone->g_nodes.list[currentNodeID].deletion_edges.length; i++) { total_count = total_count + backbone->g_nodes.list[currentNodeID].deletion_edges.list[i].weight; if (backbone->g_nodes.list[currentNodeID].deletion_edges.list[i].weight > max_count) { max_count = backbone->g_nodes.list[currentNodeID].deletion_edges.list[i].weight; max_edge = i; max_type = DELETION; } } ///这种情况下矫正 if(max_count >= total_count*CORRECT_THRESHOLD) { currentNodeID = add_path_to_correct_read(backbone, dumy, currentNodeID, max_type, max_edge, current_cigar, self_string); } else ///不矫正, 直接取下一个backbone节点 { currentNodeID++; add_base_to_correct_read_directly(dumy, backbone->g_nodes.list[currentNodeID].base); add_cigar_record(&(backbone->g_nodes.list[currentNodeID].base), 1, current_cigar, 0); } ///fprintf(stderr, "currentNodeID: %d, max_type: %d\n", currentNodeID, max_type); } else ///非backbone节点就会出错了 { fprintf(stderr, "error\n"); } } } /** ///从backbone_start遍历到backbone_end节点,生成出来的seq要接着放到dumy->corrected_read中 void get_seq_from_Graph(Graph* backbone, long long backbone_start, long long backbone_end, Correct_dumy* dumy) { long long new_seq_length = 0; long long currentNodeID; long long i; // 总共有以下几种情况: // 1. match 2. mismatch (A, C, G, T, N) 3. deletion 4. insertion (A, C, G, T) // 其实就是 1. 自己本身的weight 2. alignToNode的weight 3. insertion节点的weight long long max_count; int max_type; long long max_node; long long total_count; long long nodeID; char current_base; currentNodeID = backbone_start; while (currentNodeID != backbone_end) { total_count = 0; max_count = -1; ///图上能够被遍历到的有两种节点 ///1. backbone节点 2. insertion节点 ///backbone节点才有match/mismatch/deletion ///insertion这些都没有,就是无脑看出边 ///假如这是个backbone节点 if (currentNodeID >= backbone_start && currentNodeID <= backbone_end) { ///match total_count += backbone->g_nodes.list[currentNodeID].weight; max_count = backbone->g_nodes.list[currentNodeID].weight; max_type = 0; max_node = currentNodeID; ///mismatch和deletion (A, C, G, T, N, D, 除了自己的那个字符) for (i = 0; i < backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.length; i++) { nodeID = backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 1; max_node = nodeID; } } ///insertion (A, C, G, T, N) ///注意这个出边还得避开下一个backbone节点 for (i = 0; i < backbone->g_nodes.list[currentNodeID].outcome_edges.length; i++) { if (backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].weight == 2) { nodeID = backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 2; max_node = nodeID; } ///拿到的nodeID应该一定不是backbone上的,如果是就错了 if (nodeID >= backbone_start && nodeID <= backbone_end) { fprintf(stderr, "error\n"); } } } } else ///如果是insertion节点,就无脑看出边 { ///insertion (A, C, G, T, N) ///注意这个出边不用避开下一个backbone节点 for (i = 0; i < backbone->g_nodes.list[currentNodeID].outcome_edges.length; i++) { if (backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].weight == 2) { nodeID = backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 2; max_node = nodeID; } } else ///如果边不是2就不对了 { fprintf(stderr, "error\n"); } } } if(max_count >= total_count*CORRECT_THRESHOLD) { current_base = backbone->g_nodes.list[max_node].base; ///说明是insertion if (max_type == 2) { currentNodeID = max_node; } else ///其他情况依然沿着backbone向前 { currentNodeID++; } } else { ///假如这是个backbone节点, 不矫正 if (currentNodeID >= backbone_start && currentNodeID <= backbone_end) { current_base = backbone->g_nodes.list[currentNodeID].base; currentNodeID++; } else///如果在insertion节点上不达标很麻烦...,只能选最大的了 { current_base = backbone->g_nodes.list[max_node].base; ///说明是insertion if (max_type == 2) { currentNodeID = max_node; } else ///insertion节点不可能出现这种情况 { fprintf(stderr, "error\n"); } } } if (max_count <= 0) { fprintf(stderr, "error\n"); } add_base_to_correct_read(dumy, current_base, max_type); } ///最后还要处理backbone_end这个节点 total_count = 0; max_count = -1; ///这个节点肯定是backbone上的节点啊 ///match total_count += backbone->g_nodes.list[currentNodeID].weight; max_count = backbone->g_nodes.list[currentNodeID].weight; max_type = 0; max_node = currentNodeID; ///mismatch和deletion (A, C, G, T, N, D, 除了自己的那个字符) for (i = 0; i < backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.length; i++) { nodeID = backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 1; max_node = nodeID; } } ///这个节点不应该有任何出边了 if(backbone->g_nodes.list[currentNodeID].outcome_edges.length) { fprintf(stderr, "haha\n"); } if(max_count >= total_count*CORRECT_THRESHOLD) { current_base = backbone->g_nodes.list[max_node].base; } else { current_base = backbone->g_nodes.list[currentNodeID].base; } if (max_count <= 0) { fprintf(stderr, "error\n"); } add_base_to_correct_read(dumy, current_base, max_type); } **/ /** ///从backbone_start遍历到backbone_end节点,生成出来的seq要接着放到dumy->corrected_read中 void get_seq_from_Graph_Len2(Graph* backbone, long long backbone_start, long long backbone_end, Correct_dumy* dumy) { long long new_seq_length = 0; long long currentNodeID; long long i; // 总共有以下几种情况: // 1. match 2. mismatch (A, C, G, T, N) 3. deletion 4. insertion (A, C, G, T) // 其实就是 1. 自己本身的weight 2. alignToNode的weight 3. insertion节点的weight long long max_count; int max_type; long long max_node; long long total_count; long long nodeID; char current_base; char buffer[2]; currentNodeID = backbone_start; while (currentNodeID != backbone_end) { total_count = 0; max_count = -1; ///图上能够被遍历到的有两种节点 ///1. backbone节点 2. insertion节点 ///backbone节点才有match/mismatch/deletion ///insertion这些都没有,就是无脑看出边 ///假如这是个backbone节点 if (currentNodeID >= backbone_start && currentNodeID <= backbone_end) { ///match total_count += backbone->g_nodes.list[currentNodeID].weight; max_count = backbone->g_nodes.list[currentNodeID].weight; max_type = 0; max_node = currentNodeID; ///mismatch和deletion (A, C, G, T, N, D, 除了自己的那个字符) for (i = 0; i < backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.length; i++) { nodeID = backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 1; max_node = nodeID; } } ///insertion (A, C, G, T, N) ///注意这个出边还得避开下一个backbone节点 for (i = 0; i < backbone->g_nodes.list[currentNodeID].outcome_edges.length; i++) { if (backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].weight == 2) { ///fprintf(stderr, "error\n"); nodeID = backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 2; max_node = nodeID; } ///拿到的nodeID应该一定不是backbone上的,如果是就错了 if (nodeID >= backbone_start && nodeID <= backbone_end) { fprintf(stderr, "error\n"); } } } } else ///如果是insertion节点,就无脑看出边 { if (backbone->g_nodes.list[currentNodeID].outcome_edges.length!=1) { fprintf(stderr, "error000\n"); } ///insertion (A, C, G, T, N) ///注意这个出边不用避开下一个backbone节点 for (i = 0; i < backbone->g_nodes.list[currentNodeID].outcome_edges.length; i++) { if (backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].weight == 2) { nodeID = backbone->g_nodes.list[currentNodeID].outcome_edges.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 2; max_node = nodeID; } } else ///如果边不是2就不对了 { fprintf(stderr, "error\n"); } } } if(max_count >= total_count*CORRECT_THRESHOLD) { current_base = backbone->g_nodes.list[max_node].base; ///说明是insertion if (max_type == 2) { currentNodeID = max_node; } else ///其他情况依然沿着backbone向前 { currentNodeID++; } } else { ///假如这是个backbone节点, 不矫正 if (currentNodeID >= backbone_start && currentNodeID <= backbone_end) { current_base = backbone->g_nodes.list[currentNodeID].base; currentNodeID++; } ///应该不存在这个问题 else///如果在insertion节点上不达标很麻烦...,只能选最大的了 { ///因为现在每个insert节点只有一个出边,且这个出边到backbone fprintf(stderr, "error111\n"); } } if (max_count <= 0) { fprintf(stderr, "error\n"); } if (current_base < 'A') { buffer[0] = s_H[(current_base >> 2) & ((uint8_t)3)]; buffer[1] = s_H[current_base & ((uint8_t)3)]; add_base_to_correct_read(dumy, buffer[0], max_type); add_base_to_correct_read(dumy, buffer[1], max_type); } else { add_base_to_correct_read(dumy, current_base, max_type); } } ///最后还要处理backbone_end这个节点 total_count = 0; max_count = -1; ///这个节点肯定是backbone上的节点啊 ///match total_count += backbone->g_nodes.list[currentNodeID].weight; max_count = backbone->g_nodes.list[currentNodeID].weight; max_type = 0; max_node = currentNodeID; ///mismatch和deletion (A, C, G, T, N, D, 除了自己的那个字符) for (i = 0; i < backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.length; i++) { nodeID = backbone->g_nodes.list[currentNodeID].alignedTo_Nodes.list[i].out_node; total_count += backbone->g_nodes.list[nodeID].weight; if (backbone->g_nodes.list[nodeID].weight > max_count) { max_count = backbone->g_nodes.list[nodeID].weight; max_type = 1; max_node = nodeID; } } ///这个节点不应该有任何出边了 if(backbone->g_nodes.list[currentNodeID].outcome_edges.length) { fprintf(stderr, "haha\n"); } if(max_count >= total_count*CORRECT_THRESHOLD) { current_base = backbone->g_nodes.list[max_node].base; } else { current_base = backbone->g_nodes.list[currentNodeID].base; } if (max_count <= 0) { fprintf(stderr, "error\n"); } add_base_to_correct_read(dumy, current_base, max_type); } **/ void window_consensus(char* r_string, long long window_start, long long window_end, overlap_region_alloc* overlap_list, Correct_dumy* dumy, All_reads* R_INF, Graph* g, Cigar_record* current_cigar) { clear_Graph(g); long long x_start; long long x_length; char* x_string; char* y_string; char* backbone; long long backbone_length; long long i; long long y_start, y_length; long long overlapID, windowID; long long startNodeID, endNodeID, currentNodeID; ///这个和前面算alignment还不一样 ///那个时候x_start和x_end是当前窗口内的overlap的起始和结束位置 ///这个window就是要做consensus啊,所以起始和结束就是window本身,固定的 backbone = r_string + window_start; backbone_length = window_end - window_start + 1; addUnmatchedSeqToGraph(g, backbone, backbone_length, &startNodeID, &endNodeID); long long correct_x_pos_s; ///与当前window重叠的所有overlap for (i = 0; i < dumy->length; i++) { ///这个是那个overlap的ID,而不是overlap里对应窗口的ID overlapID = dumy->overlapID[i]; correct_x_pos_s = (overlap_list->list[overlapID].x_pos_s / WINDOW) * WINDOW; windowID = (window_start - correct_x_pos_s) / WINDOW; ///如果这个window不匹配,跳过 if (overlap_list->list[overlapID].w_list[windowID].y_end == -1) { continue; } x_start = overlap_list->list[overlapID].w_list[windowID].x_start; x_length = overlap_list->list[overlapID].w_list[windowID].x_end - overlap_list->list[overlapID].w_list[windowID].x_start + 1; y_start = overlap_list->list[overlapID].w_list[windowID].y_start; y_length = overlap_list->list[overlapID].w_list[windowID].y_end - overlap_list->list[overlapID].w_list[windowID].y_start + 1; /** fprintf(stderr, "****window_start: %d, x_start: %d, x_end: %d, x_length: %d, y_start: %d, y_length: %d, dumy->last_boundary_length: %d\n", window_start, x_start, overlap_list->list[overlapID].w_list[windowID].x_end , x_length, y_start, y_length, dumy->last_boundary_length); **/ recover_UC_Read_sub_region(dumy->overlap_region, y_start, y_length, overlap_list->list[overlapID].y_pos_strand, R_INF, overlap_list->list[overlapID].y_id); x_string = r_string + x_start; y_string = dumy->overlap_region; ///这个是比对上的起始base在backbone上对应的位置,也就是节点ID currentNodeID = x_start - window_start; ///这个是要用的cigar: overlap_list->list[overlapID].w_list[windowID].cigar; addmatchedSeqToGraph(g, currentNodeID, x_string, x_length, y_string, y_length, &(overlap_list->list[overlapID].w_list[windowID].cigar), startNodeID, endNodeID); } get_seq_from_Graph(g, dumy, current_cigar, backbone); ///get_seq_from_Graph(g, startNodeID, endNodeID, dumy); ///get_seq_from_Graph_Len2(g, startNodeID, endNodeID, dumy); ///debug_graph(g, backbone_length); /** for (i = 0; i < dumy->length; i++) { ///这个是那个overlap的ID,而不是overlap里对应窗口的ID overlapID = dumy->overlapID[i]; correct_x_pos_s = (overlap_list->list[overlapID].x_pos_s / WINDOW) * WINDOW; windowID = (window_start - correct_x_pos_s) / WINDOW; ///如果这个window不匹配,跳过 if (overlap_list->list[overlapID].w_list[windowID].y_end == -1) { continue; } x_start = overlap_list->list[overlapID].w_list[windowID].x_start; x_length = overlap_list->list[overlapID].w_list[windowID].x_end - overlap_list->list[overlapID].w_list[windowID].x_start + 1; y_start = overlap_list->list[overlapID].w_list[windowID].y_start; y_length = overlap_list->list[overlapID].w_list[windowID].y_end - overlap_list->list[overlapID].w_list[windowID].y_start + 1; recover_UC_Read_sub_region(dumy->overlap_region, y_start, y_length, overlap_list->list[overlapID].y_pos_strand, R_INF, overlap_list->list[overlapID].y_id); x_string = r_string + x_start; y_string = dumy->overlap_region; ///这个是比对上的起始base在backbone上对应的位置,也就是节点ID currentNodeID = x_start - window_start; ///这个是要用的cigar: overlap_list->list[overlapID].w_list[windowID].cigar; Graph_debug(g, currentNodeID, x_string, x_length, y_string, y_length, &(overlap_list->list[overlapID].w_list[windowID].cigar), startNodeID, endNodeID); } for (i = 0; i < g->g_nodes.length; i++) { if (i >= startNodeID && i <= endNodeID) { if (g->g_nodes.list[i].weight != 1) { fprintf(stderr, "error 1\n"); } } else { if (g->g_nodes.list[i].weight != 0) { fprintf(stderr, "error 2\n"); } if (g->g_nodes.list[i].alignedTo_Nodes.length != 0) { fprintf(stderr, "error 3\n"); } ///节点入边不为0,说明这个不是alignTO节点,而是insert节点 if (g->g_nodes.list[i].income_edges.length != 0 && g->g_nodes.list[i].outcome_edges.length != 0) { fprintf(stderr, "error 4\n"); } } } **/ } void add_cigar_to_cigar(Correct_dumy* backbone_dumy, Cigar_record* backbone_cigar, Round2_alignment* second_round, long long back_bone_start, long long back_bone_length, long long new_start, long long new_length) { Correct_dumy* new_dumy = &(second_round->dumy); Cigar_record* new_cigar = &(second_round->tmp_cigar); Cigar_record* result_cigar = &(second_round->cigar); char* x_string = backbone_dumy->corrected_read + back_bone_start; char* y_string = new_dumy->corrected_read + new_start; /** if(verify_cigar_2(x_string, back_bone_length, y_string, new_length, new_cigar, -1)) { fprintf(stderr, "error\n"); } **/ ///if type == 0, x_string here is not useful ///output matches to cigar add_cigar_record(x_string, back_bone_start - second_round->obtained_cigar_length, result_cigar, 0); second_round->obtained_cigar_length = back_bone_start + back_bone_length; long long i, cigar_i, x_i, y_i; int operation; int operationLen; x_i = y_i = 0; char merge_base; for (i = 0; i < new_cigar->length; i++) { operation = Get_Cigar_Type(new_cigar->record[i]); operationLen = Get_Cigar_Length(new_cigar->record[i]); if (operation == 0) { ///if type == 0, x_string here is not useful add_cigar_record(x_string, operationLen, result_cigar, 0); x_i += operationLen; y_i += operationLen; } else if (operation == 1) { for (cigar_i = 0; cigar_i < operationLen; cigar_i++) { merge_base = x_string[x_i]; merge_base = merge_base << 3; merge_base = merge_base | y_string[y_i]; add_cigar_record(&merge_base, 1, result_cigar, 1); x_i++; y_i++; } } else if (operation == INSERTION)///2是x缺字符(y多字符) { add_cigar_record(y_string+y_i, operationLen, result_cigar, INSERTION); y_i += operationLen; } else if (operation == DELETION) { add_cigar_record(x_string+x_i, operationLen, result_cigar, DELETION); x_i += operationLen; } } } ///correct bases of current_dumy->corrected_read in [start_base, end_base] int merge_cigars(Correct_dumy* current_dumy, Cigar_record* current_cigar, Round2_alignment* second_round, long long total_start_base, long long total_end_base, long long total_window_start, long long total_window_end) { Cigar_record* new_cigar = &(second_round->tmp_cigar); Correct_dumy* new_dumy = &(second_round->dumy); if(new_cigar->length == 1 && Get_Cigar_Type(new_cigar->record[0]) == 0) { return 1; } long long start_base = total_start_base - total_window_start; long long end_base = total_end_base - total_window_start; long long x_i, y_i, cigar_i, i; x_i = 0; y_i = 0; int operation; int operationLen; int cigar_error = 0; long long get_x_start, get_x_end, get_y_start, get_y_end; get_x_start = get_x_end = get_y_start = get_y_end = -1; int start_cigar = -1; int end_cigar = -1; char merge_base; ///0 is match, 1 is mismatch, 2 is up, 3 is left ///2是x缺字符(y多字符),而3是y缺字符(x多字符) ///obtained x_i may larger than start_base/end_base ///when operation == 3 ///so for operation == 3, we need deal with carefully for (i = 0; i < new_cigar->length; i++) { operation = Get_Cigar_Type(new_cigar->record[i]); operationLen = Get_Cigar_Length(new_cigar->record[i]); if (operation == 0) { for (cigar_i = 0; cigar_i < operationLen; cigar_i++) { if(x_i >= start_base && get_x_start == -1) { get_x_start = x_i; get_y_start = y_i; start_cigar = i; } if(x_i >= end_base && get_x_end == -1) { get_x_end = x_i; get_y_end = y_i; end_cigar = i; break; } x_i++; y_i++; } } else if (operation == 1) { for (cigar_i = 0; cigar_i < operationLen; cigar_i++) { if(x_i >= start_base && get_x_start == -1) { get_x_start = x_i; get_y_start = y_i; start_cigar = i; } if(x_i >= end_base && get_x_end == -1) { get_x_end = x_i; get_y_end = y_i; end_cigar = i; break; } x_i++; y_i++; } }///2是x缺字符(y多字符) else if (operation == 2) { y_i += operationLen; }///3是y缺字符(x多字符) else if (operation == 3) { ///obtained x_i may larger than start_base/end_base ///when operation == 3 ///so for operation == 3, we need deal with carefully x_i += operationLen; } } ///if there are some gap at the end of x, it very likely miscorrection if(get_x_end == -1 || get_x_start == -1) { return 0; } /** fprintf(stderr, "get_x_start: %d, get_x_end: %d\n", get_x_start, get_x_end); fprintf(stderr, "get_y_start: %d, get_y_end: %d\n", get_y_start, get_y_end); for (int ijk = 0; ijk < new_cigar->length; ijk++) { fprintf(stderr, "Oper: %d, Len: %d\n", Get_Cigar_Type(new_cigar->record[ijk]), Get_Cigar_Length(new_cigar->record[ijk])); } **/ x_i = 0; y_i = 0; uint32_t single_record = 0; for (i = 0; i < new_cigar->length; i++) { operation = Get_Cigar_Type(new_cigar->record[i]); operationLen = Get_Cigar_Length(new_cigar->record[i]); if (i == start_cigar) { single_record = 0; single_record = operationLen - (get_x_start - x_i); single_record = single_record << 2; single_record = single_record | operation; new_cigar->record[i] = single_record; if(operation > 1) { fprintf(stderr, "error\n"); } if (i == end_cigar) { x_i = get_x_start; single_record = 0; single_record = get_x_end - x_i + 1; single_record = single_record << 2; single_record = single_record | operation; new_cigar->record[i] = single_record; if(operation > 1) { fprintf(stderr, "error\n"); } break; } } else if (i == end_cigar) { single_record = 0; single_record = get_x_end - x_i + 1; single_record = single_record << 2; single_record = single_record | operation; new_cigar->record[i] = single_record; if(operation > 1) { fprintf(stderr, "error\n"); } break; } if (operation == 0 || operation == 1) { x_i += operationLen; y_i += operationLen; } else if (operation == 2) { y_i += operationLen; }///3是y缺字符(x多字符) else if (operation == 3) { x_i += operationLen; } } new_cigar->length = end_cigar - start_cigar + 1; ///可以优化 memmove(new_cigar->record, new_cigar->record + start_cigar, new_cigar->length*sizeof(uint32_t)); /** fprintf(stderr, "start_cigar: %d, end_cigar: %d\n", start_cigar, end_cigar); for (int ijk = 0; ijk < new_cigar->length; ijk++) { fprintf(stderr, "new:Oper: %d, Len: %d\n", Get_Cigar_Type(new_cigar->record[ijk]), Get_Cigar_Length(new_cigar->record[ijk])); } **/ long long total_x_start = total_window_start + get_x_start; long long x_length = get_x_end -get_x_start + 1; long long total_y_start = get_y_start; long long y_length = get_y_end -get_y_start + 1; add_cigar_to_cigar(current_dumy, current_cigar, second_round, total_x_start, x_length, total_y_start, y_length); } int process_boundary(overlap_region_alloc* overlap_list, All_reads* R_INF, Correct_dumy* dumy, Graph* g, Cigar_record* current_cigar, long long uncorrected_window_start, Round2_alignment* second_round) { char* r_string = dumy->corrected_read; long long corrected_window_start, corrected_window_end; int extra_begin; int extra_end; if(dumy->last_boundary_length == 0) { return 0; } corrected_window_start = dumy->last_boundary_length - WINDOW_BOUNDARY/2; corrected_window_end = dumy->last_boundary_length + WINDOW_BOUNDARY/2 - 1; if(corrected_window_start < 0) { corrected_window_start = 0; } if (corrected_window_end >= current_cigar->new_read_length) { corrected_window_end = current_cigar->new_read_length - 1; } clear_Graph(g); long long x_start, x_end; long long x_length, x_len, o_len; int threshold; long long Window_Len; char* x_string; char* y_string; char* backbone; long long backbone_length; long long i; long long y_start, y_length; long long overlapID, windowID; long long startNodeID, endNodeID, currentNodeID; int end_site; unsigned int error; int real_y_start; window_list tmp_cigar; long long total_error = 0; backbone = r_string + corrected_window_start; backbone_length = corrected_window_end - corrected_window_start + 1; addUnmatchedSeqToGraph(g, backbone, backbone_length, &startNodeID, &endNodeID); long long correct_x_pos_s; long long matched_coverage = 0; for (i = 0; i < dumy->length; i++) { overlapID = dumy->overlapID[i]; correct_x_pos_s = (overlap_list->list[overlapID].x_pos_s / WINDOW) * WINDOW; windowID = (uncorrected_window_start - correct_x_pos_s) / WINDOW; ///如果这个window不匹配,跳过 if (overlap_list->list[overlapID].w_list[windowID].y_end == -1) { continue; } x_start = overlap_list->list[overlapID].w_list[windowID].x_start; y_start = overlap_list->list[overlapID].w_list[windowID].y_start; /** * There are total 3 cases: * 1. this window of x is overlapped totally by y * x: ------|------|--------- * y: ------|------|--------- * in this case, x_start == uncorrected_window_start, x_length == WINDOW * 2. the suiffx of x's window is overlapped by the prefix of y * x: ------|------|--------- * y: |--|----------- * in this case, x_start > uncorrected_window_start, x_length < WINDOW * this overlap is useless * 3. the prefix of x's window is overlapped by y (see last window) * x: |------|------|-----|--- * y: |--|-----|------ * or * x: |------|------|-----|---- * y: --|------|------|-----|-- * * in this case, x_start == uncorrected_window_start, x_length < WINDOW * * case 1 and case 3 are useful, while case 2 is useless * **/ ///case 1 and case 3 are useful if(x_start == uncorrected_window_start) { extra_begin = extra_end = 0; x_start = corrected_window_start; x_end = corrected_window_end; x_len = x_end - x_start + 1; threshold = x_len * THRESHOLD_RATE; ///y_start may less than 0 y_start = y_start - WINDOW_BOUNDARY/2; ///其实可以不加...怕出bug if(y_start < 0) { continue; } Window_Len = x_len + (threshold << 1); determine_overlap_region(threshold, y_start, overlap_list->list[overlapID].y_id, Window_Len, R_INF, &extra_begin, &extra_end, &y_start, &o_len); fill_subregion(dumy->overlap_region, y_start, o_len, overlap_list->list[overlapID].y_pos_strand, R_INF, overlap_list->list[overlapID].y_id, extra_begin, extra_end); x_string = r_string + x_start; y_string = dumy->overlap_region; ///both end site and real_y_start have extra_begin ///有很多是完全匹配,可以先快速判断是不是完全匹配 end_site = Reserve_Banded_BPM_PATH(y_string, Window_Len, x_string, x_len, threshold, &error, &real_y_start, &(dumy->path_length), dumy->matrix_bit, dumy->path, -1, -1); if (error!=(unsigned int)-1) { total_error = total_error + error; matched_coverage++; tmp_cigar.x_start = x_start; tmp_cigar.x_end = x_end; generate_cigar(dumy->path, dumy->path_length, &tmp_cigar, &real_y_start, &end_site, error); ///both end site and real_y_start have extra_begin real_y_start -= extra_begin; end_site -= extra_begin; y_length = end_site - real_y_start + 1; y_start = y_start + real_y_start; x_start = corrected_window_start; x_length = corrected_window_end - x_start + 1; ///here can be improved, make y_string = dumy->overlap_region + real_y_start + extra_begin recover_UC_Read_sub_region(dumy->overlap_region, y_start, y_length, overlap_list->list[overlapID].y_pos_strand, R_INF, overlap_list->list[overlapID].y_id); x_string = r_string + x_start; y_string = dumy->overlap_region; /** if(memcmp("m54238_180914_183539/66650355/ccs", Get_NAME((*R_INF), overlap_list->list[overlapID].x_id), Get_NAME_LENGTH((*R_INF), overlap_list->list[overlapID].x_id)) == 0 && uncorrected_window_start == 375) { fprintf(stderr, "###### %.*s, x_string[124]: %c, y_string[124]: %c\n", Get_NAME_LENGTH((*R_INF), overlap_list->list[overlapID].y_id), Get_NAME((*R_INF), overlap_list->list[overlapID].y_id), x_string[124], y_string[124]); fprintf(stderr, "######error: %d****\n", error); for (int ijk = 0; ijk < tmp_cigar.cigar.length; ijk++) { fprintf(stderr, "###### Oper: %d, Len: %d\n", tmp_cigar.cigar.C_C[ijk], tmp_cigar.cigar.C_L[ijk]); } fprintf(stderr, "######dumy->path_length: %d\n****\n", dumy->path_length); } **/ /** if(verify_cigar(x_string, x_length, y_string, y_length, &tmp_cigar.cigar, error)) { fprintf(stderr, "*******error: %d****\n", error); for (int ijk = 0; ijk < tmp_cigar.cigar.length; ijk++) { fprintf(stderr, "Oper: %d, Len: %d\n", tmp_cigar.cigar.C_C[ijk], tmp_cigar.cigar.C_L[ijk]); } fprintf(stderr, "*******dumy->path_length: %d\n****\n", dumy->path_length); } **/ currentNodeID = x_start - corrected_window_start; addmatchedSeqToGraph(g, currentNodeID, x_string, x_length, y_string, y_length, &(tmp_cigar.cigar), startNodeID, endNodeID); } }///case 2 is useless else if(x_start != uncorrected_window_start) { continue; } } /** fprintf(stderr, "matched_coverage: %d, dumy->length: %d\n", matched_coverage, dumy->length); **/ if(matched_coverage >= MIN_COVERAGE_THRESHOLD) { ///if there are no error, we do not need correction if(total_error == 0) { return 0; } /** fprintf(stderr, "s_start_nodeID: %d, s_end_nodeID: %d, corrected_window_start: %d, corrected_window_end: %d\n", g->s_start_nodeID, g->s_end_nodeID, corrected_window_start, corrected_window_end); **/ clear_Cigar_record(&(second_round->tmp_cigar)); clear_Correct_dumy_pure(&(second_round->dumy)); ///correct bases in [start_base, end_base] long long start_base = corrected_window_start + WINDOW_UNCORRECT_SINGLE_SIDE_BOUNDARY; long long end_base = corrected_window_end - WINDOW_UNCORRECT_SINGLE_SIDE_BOUNDARY; if(end_base > start_base) { ///note there is an additional "S" node ///and start from i-th node, we can correct (i+1)-th base /// so the condition when traversing graph is ///(node >= start_base - corrected_window_start && node <= end_base - corrected_window_start) get_seq_from_Graph(g, &(second_round->dumy), &(second_round->tmp_cigar), backbone); /** if(verify_cigar_2(backbone, backbone_length, second_round->dumy.corrected_read, second_round->dumy.corrected_read_length, &(second_round->tmp_cigar), -1)) { fprintf(stderr, "hahah\n"); } **/ merge_cigars(dumy, current_cigar, second_round, start_base, end_base, corrected_window_start, corrected_window_end); } } else { return 0; } return 1; } void generate_consensus(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, Correct_dumy* dumy, Graph* g, Cigar_record* current_cigar, Round2_alignment* second_round) { clear_Cigar_record(current_cigar); long long window_num = (g_read->length + WINDOW - 1) / WINDOW; long long i, j, overlap_length; long long window_start, window_end; long long num_availiable_win = 0; window_start = 0; window_end = WINDOW - 1; if (window_end >= g_read->length) { window_end = g_read->length - 1; } int flag; ///for last window dumy->last_boundary_length = 0; for (i = 0; i < window_num; i++) { dumy->length = 0; dumy->lengthNT = 0; ///flag返回的是重叠数量 ///dumy->length返回的是有效完全重叠的数量 ///dumy->lengthNT返回的是有效不完全重叠的数量 ///return overlaps that is overlaped with [window_start, window_end] flag = get_available_interval(window_start, window_end, overlap_list, dumy); switch (flag) { case 1: ///找到匹配 break; case 0: ///没找到匹配 break; case -2: ///下一个window也不会存在匹配, 直接跳出 i = window_num; break; } ///这个是available overlap里所有window的数量... ///num_availiable_win = num_availiable_win + dumy->length + dumy->lengthNT; num_availiable_win = num_availiable_win + dumy->length; ///重叠窗口数,也就是coverage大小 if(dumy->length >= MIN_COVERAGE_THRESHOLD) { window_consensus(g_read->seq, window_start, window_end, overlap_list, dumy, R_INF, g, current_cigar); if(dumy->last_boundary_length != 0) { process_boundary(overlap_list, R_INF, dumy, g, current_cigar, window_start, second_round); /** if(memcmp("m54238_180914_183539/66650355/ccs", Get_NAME((*R_INF), overlap_list->list[0].x_id), Get_NAME_LENGTH((*R_INF), overlap_list->list[0].x_id)) == 0) { ///fprintf(stderr, "%.*s\n\n", Get_NAME_LENGTH((*R_INF), overlap_list->list[0].x_id), Get_NAME((*R_INF), overlap_list->list[0].x_id)); fprintf(stderr, "window_start: %d, window_end: %d, dumy->last_boundary_length: %d\n", window_start, window_end, dumy->last_boundary_length); fprintf(stderr, "new_read_length: %d\n",second_round->cigar.new_read_length); fprintf(stderr, "cigar.length: %d\n",second_round->cigar.length); fprintf(stderr, "obtained_cigar_length: %d\n",second_round->obtained_cigar_length); fprintf(stderr, "*******\n"); for (size_t ijk = 0; ijk < second_round->cigar.length; ijk++) { int operation = Get_Cigar_Type(second_round->cigar.record[ijk]); int operation_length = Get_Cigar_Length(second_round->cigar.record[ijk]); fprintf(stderr, "oper: %d, oper_len: %d\n", operation, operation_length); } fprintf(stderr, "*******\n"); } **/ } } else { add_segment_to_correct_read(dumy, g_read->seq + window_start, window_end - window_start + 1); add_cigar_record(g_read->seq + window_start, window_end - window_start + 1, current_cigar, 0); } dumy->last_boundary_length = current_cigar->new_read_length; window_start = window_start + WINDOW; window_end = window_end + WINDOW; if (window_end >= g_read->length) { window_end = g_read->length - 1; } } if (window_start < g_read->length) { add_segment_to_correct_read(dumy, g_read->seq + window_start, g_read->length - window_start); add_cigar_record(g_read->seq + window_start, g_read->length - window_start, current_cigar, 0); } ///if type == 0, x_string here is not useful ///output matches to cigar if (current_cigar->new_read_length != second_round->obtained_cigar_length) { add_cigar_record(dumy->corrected_read, current_cigar->new_read_length - second_round->obtained_cigar_length, &(second_round->cigar), 0); } } void markSNP( long long window_offset, long long x_total_start, long long x_length, long long y_total_start, long long y_length, CIGAR* cigar, haplotype_evdience_alloc* hap) { int x_i, y_i, cigar_i; x_i = 0; y_i = 0; cigar_i = 0; int operation; int operationLen; int i; long long inner_offset = x_total_start - window_offset; ///note that node 0 is the start node ///0 is match, 1 is mismatch, 2 is up, 3 is left ///2是x缺字符(y多字符),而3是y缺字符(x多字符) while (cigar_i < cigar->length) { operation = cigar->C_C[cigar_i]; operationLen = cigar->C_L[cigar_i]; ///这种情况代表匹配和mismatch if (operation == 0) { x_i += operationLen; y_i += operationLen; } else if(operation == 1) { for (i = 0; i < operationLen; i++) { /** if(inner_offset + x_i >= WINDOW) { fprintf(stderr, "error\n"); } **/ hap->flag[inner_offset + x_i]++; x_i++; y_i++; } }///insertion else if (operation == 2) { y_i += operationLen; } else if (operation == 3) { x_i += operationLen; } cigar_i++; } /** if(x_i != x_length || y_i != y_length) { fprintf(stderr, "x_i: %d, x_length: %d\n", x_i, x_length); fprintf(stderr, "y_i: %d, y_length: %d\n", y_i, y_length); } int no_zero = 0; for (i = 0; i < WINDOW; i++) { if(hap->flag[i] != 0) { no_zero++; } } fprintf(stderr, "no_zero: %d\n", no_zero); **/ } void addSNPtohaplotype( long long window_offset, int overlapID, char* x_string, long long x_total_start, long long x_length, char* y_string, long long y_total_start, long long y_length, CIGAR* cigar, haplotype_evdience_alloc* hap) { int x_i, y_i, cigar_i; x_i = 0; y_i = 0; cigar_i = 0; int operation; int operationLen; int i; long long inner_offset = x_total_start - window_offset; haplotype_evdience ev; ///note that node 0 is the start node ///0 is match, 1 is mismatch, 2 is up, 3 is left ///2是x缺字符(y多字符),而3是y缺字符(x多字符) while (cigar_i < cigar->length) { operation = cigar->C_C[cigar_i]; operationLen = cigar->C_L[cigar_i]; ///这种情况代表匹配和mismatch if (operation == 0) { for (i = 0; i < operationLen; i++) { if(hap->flag[inner_offset] > FLAG_THRE) { ev.misBase = y_string[y_i]; ev.overlapID = overlapID; ev.site = x_total_start + x_i; ev.overlapSite = y_total_start + y_i; ev.type = 0; addHaplotypeEvdience(hap, &ev); } inner_offset++; x_i++; y_i++; } } else if(operation == 1) { for (i = 0; i < operationLen; i++) { if(hap->flag[inner_offset] > FLAG_THRE) { ev.misBase = y_string[y_i]; ev.overlapID = overlapID; ev.site = x_total_start + x_i; ev.overlapSite = y_total_start + y_i; ev.type = 1; addHaplotypeEvdience(hap, &ev); } inner_offset++; x_i++; y_i++; } }///insertion else if (operation == 2) { y_i += operationLen; } else if (operation == 3) { for (i = 0; i < operationLen; i++) { if(hap->flag[inner_offset] > FLAG_THRE) { ev.misBase = 'N'; ev.overlapID = overlapID; ev.site = x_total_start + x_i; ev.overlapSite = y_total_start + y_i; ev.type = 2; addHaplotypeEvdience(hap, &ev); } inner_offset++; x_i++; } } cigar_i++; } } void cluster(char* r_string, long long window_start, long long window_end, overlap_region_alloc* overlap_list, Correct_dumy* dumy, All_reads* R_INF, haplotype_evdience_alloc* hap) { long long x_start; long long x_length; char* x_string; char* y_string; long long i; long long y_start, y_length; long long overlapID, windowID; long long startNodeID, endNodeID, currentNodeID; RsetInitHaplotypeEvdienceFlag(hap); long long correct_x_pos_s; long long inner_window_offset; ///与当前window重叠的所有overlap ///first mark all snp pos for (i = 0; i < dumy->length; i++) { ///这个是那个overlap的ID,而不是overlap里对应窗口的ID overlapID = dumy->overlapID[i]; ///overlap_list->list[overlapID].x_pos_s is the begining of the whole overlap correct_x_pos_s = (overlap_list->list[overlapID].x_pos_s / WINDOW) * WINDOW; ///window_start is the begining of this window in the whole x_read windowID = (window_start - correct_x_pos_s) / WINDOW; ///如果这个window不匹配,跳过 if (overlap_list->list[overlapID].w_list[windowID].y_end == -1) { continue; } ///both x_start and y_start are the begining of the whole x_read and y_read x_start = overlap_list->list[overlapID].w_list[windowID].x_start; x_length = overlap_list->list[overlapID].w_list[windowID].x_end - overlap_list->list[overlapID].w_list[windowID].x_start + 1; y_start = overlap_list->list[overlapID].w_list[windowID].y_start; y_length = overlap_list->list[overlapID].w_list[windowID].y_end - overlap_list->list[overlapID].w_list[windowID].y_start + 1; markSNP(window_start, x_start, x_length, y_start, y_length, &(overlap_list->list[overlapID].w_list[windowID].cigar), hap); } for (i = 0; i < WINDOW; i++) { if(hap->flag[i] > FLAG_THRE) { hap->snp++; } } ///add the information related to snp to haplotype_evdience_alloc for (i = 0; i < dumy->length; i++) { ///这个是那个overlap的ID,而不是overlap里对应窗口的ID overlapID = dumy->overlapID[i]; ///overlap_list->list[overlapID].x_pos_s is the begining of the whole overlap correct_x_pos_s = (overlap_list->list[overlapID].x_pos_s / WINDOW) * WINDOW; ///window_start is the begining of this window in the whole x_read windowID = (window_start - correct_x_pos_s) / WINDOW; ///如果这个window不匹配,跳过 if (overlap_list->list[overlapID].w_list[windowID].y_end == -1) { continue; } ///both x_start and y_start are the begining of the whole x_read and y_read x_start = overlap_list->list[overlapID].w_list[windowID].x_start; x_length = overlap_list->list[overlapID].w_list[windowID].x_end - overlap_list->list[overlapID].w_list[windowID].x_start + 1; y_start = overlap_list->list[overlapID].w_list[windowID].y_start; y_length = overlap_list->list[overlapID].w_list[windowID].y_end - overlap_list->list[overlapID].w_list[windowID].y_start + 1; recover_UC_Read_sub_region(dumy->overlap_region, y_start, y_length, overlap_list->list[overlapID].y_pos_strand, R_INF, overlap_list->list[overlapID].y_id); x_string = r_string + x_start; y_string = dumy->overlap_region; addSNPtohaplotype(window_start, overlapID, x_string, x_start, x_length, y_string, y_start, y_length, &(overlap_list->list[overlapID].w_list[windowID].cigar), hap); } } int cmp_haplotype_evdience(const void * a, const void * b) { if ((*(haplotype_evdience*)a).site != (*(haplotype_evdience*)b).site) { return (*(haplotype_evdience*)a).site > (*(haplotype_evdience*)b).site ? 1 : -1; } else { if ((*(haplotype_evdience*)a).type != (*(haplotype_evdience*)b).type) { return (*(haplotype_evdience*)a).type > (*(haplotype_evdience*)b).type ? 1 : -1; } else { if ((*(haplotype_evdience*)a).misBase != (*(haplotype_evdience*)b).misBase) { return (*(haplotype_evdience*)a).misBase > (*(haplotype_evdience*)b).misBase ? 1 : -1; } else { return 0; } } } } int cmp_snp_stats(const void * a, const void * b) { if ((*(SnpStats*)a).score != (*(SnpStats*)b).score) { return (*(SnpStats*)a).score < (*(SnpStats*)b).score ? 1 : -1; } else { if ((*(SnpStats*)a).occ_2 != (*(SnpStats*)b).occ_2) { return (*(SnpStats*)a).occ_2 > (*(SnpStats*)b).occ_2 ? 1 : -1; } else { return 0; } } } void debug_hap_information(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, haplotype_evdience_alloc* hap, Correct_dumy* dumy) { int i, overlapID, y_ID, y_Strand; long long x_start; long long x_length; char* x_string; char* y_string; long long y_start; long long y_length; for (i = 0; i < hap->length; i++) { if(hap->list[i].type < 2) { overlapID = hap->list[i].overlapID; x_start = hap->list[i].site; y_start = hap->list[i].overlapSite; y_ID = overlap_list->list[overlapID].y_id; y_Strand = overlap_list->list[overlapID].y_pos_strand; recover_UC_Read_sub_region(dumy->overlap_region, y_start, 1, y_Strand, R_INF, y_ID); x_string = g_read->seq + x_start; y_string = dumy->overlap_region; if(y_string[0] != hap->list[i].misBase) { fprintf(stderr, "y_string[0]: %c, hap->list[i].misBase: %c\n", y_string[0], hap->list[i].misBase); } if(hap->list[i].type == 0) { if(x_string[0] != y_string[0]) { fprintf(stderr, "x_string[0]: %c, y_string[0]: %c\n", x_string[0], y_string[0]); } } else if(hap->list[i].type == 0) { if(x_string[0] == y_string[0]) { fprintf(stderr, "x_string[0]: %c, y_string[0]: %c\n", x_string[0], y_string[0]); } } } } } int debug_split_sub_list(haplotype_evdience_alloc* hap, haplotype_evdience* sub_list, long long sub_length, long long num_haplotype) { long long i = 0; long long occ_0 = 0; long long occ_1 = 0; long long occ_1_array[5]; memset(occ_1_array, 0, sizeof(long long) * 5); long long occ_2 = 0; for (i = 0; i < sub_length; i++) { if(sub_list[i].type == 0) { occ_0++; } else if(sub_list[i].type == 1) { occ_1_array[seq_nt6_table[(uint8_t)(sub_list[i].misBase)]]++; occ_1++; } else if(sub_list[i].type == 2) { occ_2++; } } /** 1. if occ_0 = 0, that means all overlaps are different with this read at this site 2. it is not possible that occ_1 = 0, 3. if occ_1 = 1, there are only one difference. It must be a sequencing error. **/ if(occ_0 == 0 || occ_1 <= 1) { return 0; } ///note: if the max value except type0 is type2 ///that means this is no snp hapolyte long long max = occ_2; long long max_i = -1; for (i = 0; i < 5; i++) { if(occ_1_array[i] > max) { max = occ_1_array[i]; max_i = i; } } if(max_i == -1) { return 0; } if(max <= 1) { return 0; } ///if we have two max for (i = 0; i < 5; i++) { if(occ_1_array[i] == max && i != max_i) { return 0; } } long long new_0 = occ_0 + 1; long long new_total = sub_length + 1; ///note: here occ_0++ since the read itself has a type0 double available = new_0 + max; double threshold = 0.95; available = available/((double)(new_total)); if(available < threshold) { return 0; } ///if we just have one snp, we need to phase it carefully if(num_haplotype == 1) { ///we must have just 1 match and 1 mismatch ///any other types are not good if(new_0 + max != new_total) { return 0; } if(filter_snp(new_0, max, new_total) == 0) { return 0; } } /** if(filter_snp(new_0, max, new_total) == 0) { return 0; } **/ ///for each calculated snp, find if it is at snp matrix for (i = 0; i < hap->available_snp; i++) { if(hap->snp_stat[i].site == sub_list[0].site) { int j = 0; int vectorID = hap->snp_stat[i].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); if(hap->snp_stat[i].occ_0 != occ_0) { fprintf(stderr, "error occ0\n"); } if(hap->snp_stat[i].occ_1 != occ_1_array[max_i]) { fprintf(stderr, "error occ1\n"); } if(hap->snp_stat[i].overlap_num != sub_length) { fprintf(stderr, "error overlap_num\n"); } if(hap->snp_stat[i].overlap_num != hap->snp_stat[i].occ_0 + hap->snp_stat[i].occ_1 + hap->snp_stat[i].occ_2) { fprintf(stderr, "error overlap_num\n"); } ///for each element in snp vector, find if it is in calculated dataset for (j = 0; j < Get_SNP_Vector_Length((*hap)); j++) { if(vector[j] != -1) { int x_i = 0; for (x_i = 0; x_i < sub_length; x_i++) { if(j == sub_list[x_i].overlapID) { break; } } if(x_i == sub_length) { fprintf(stderr, "error: j: %d\n",j); } else { if(vector[j] == 0 || sub_list[x_i].type == 0) { if(vector[j] != sub_list[x_i].type) { fprintf(stderr, "error: 0: %d\n",j); } } if(vector[j] == 1) { if(sub_list[x_i].type != 1) { fprintf(stderr, "-error: 1: %d\n",j); } if(sub_list[x_i].type == 1 && sub_list[x_i].misBase != s_H[max_i]) { fprintf(stderr, "+error: 1: %d\n",j); } } if(vector[j] == 2) { if(sub_list[x_i].type != 2) { if(sub_list[x_i].type == 1 && sub_list[x_i].misBase != s_H[max_i]) { ; } else { fprintf(stderr, "error: 2: %d\n",j); } } } } } } ///for each calculated data, find if it is in snp vector for (j = 0; j < sub_length; j++) { if(vector[sub_list[j].overlapID] != sub_list[j].type) { if(vector[sub_list[j].overlapID] == 2 && sub_list[j].type == 1 && sub_list[j].misBase != s_H[max_i]) { ; } else { fprintf(stderr, "vector[sub_list[j].site]: %d, sub_list[j].type: %d\n", vector[sub_list[j].overlapID], sub_list[j].type); } } } break; } } if(i == hap->available_snp) { fprintf(stderr, "error\n"); } /** fprintf(stderr, "new_0: %d, occ_0: %d, max: %d, max_i: %d, sub_length: %d, new_total: %d, available: %lf\n", new_0, occ_0, max, max_i, sub_length, new_total, available); for (i = 0; i < sub_length; i++) { fprintf(stderr, "i: %d, site: %d, type: %d, char: %c, ID: %d, name: %.*s\n", i, sub_list[i].site, sub_list[i].type, sub_list[i].misBase, sub_list[i].overlapID, Get_NAME_LENGTH((*R_INF), overlap_list->list[sub_list[i].overlapID].y_id), Get_NAME((*R_INF),overlap_list->list[sub_list[i].overlapID].y_id)); } fprintf(stderr, "\n"); **/ return 1; } int debug_snp_matrix(haplotype_evdience_alloc* hap) { uint64_t pre_site = (uint64_t)-1; uint64_t num_of_snps = 0; long long pre_i = -1; long long sub_length; haplotype_evdience* sub_list; long long i; long long a_snp = 0; ////split reads for (i = 0; i < hap->length; i++) { if(pre_site != hap->list[i].site) { if(i != 0) { sub_list = hap->list + pre_i; sub_length = i - pre_i; ///debug_total_length = debug_total_length + sub_length; a_snp += debug_split_sub_list(hap, sub_list, sub_length, hap->snp); } num_of_snps++; pre_site = hap->list[i].site; pre_i = i; } } if(pre_i != -1) { sub_list = hap->list + pre_i; sub_length = i - pre_i; ///debug_total_length = debug_total_length + sub_length; a_snp += debug_split_sub_list(hap, sub_list, sub_length, hap->snp); } if(a_snp != hap->available_snp) { fprintf(stderr, "a_snp: %d, available_snp: %d\n", a_snp, hap->available_snp); } } int split_sub_list(haplotype_evdience_alloc* hap, haplotype_evdience* sub_list, long long sub_length, long long num_haplotype, overlap_region_alloc* overlap_list, All_reads* R_INF) { long long i = 0; long long occ_0 = 0; long long occ_1 = 0; long long occ_1_array[5]; memset(occ_1_array, 0, sizeof(long long) * 5); long long occ_2 = 0; for (i = 0; i < sub_length; i++) { if(sub_list[i].type == 0) { occ_0++; } else if(sub_list[i].type == 1) { occ_1_array[seq_nt6_table[(uint8_t)(sub_list[i].misBase)]]++; occ_1++; } else if(sub_list[i].type == 2) { occ_2++; } } /** 1. if occ_0 = 0, that means all overlaps are different with this read at this site 2. it is not possible that occ_1 = 0, 3. if occ_1 = 1, there are only one difference. It must be a sequencing error. **/ if(occ_0 == 0 || occ_1 <= 1) { return 0; } ///note: if the max value except type0 is type2 ///that means this is no snp hapolyte long long max = occ_2; long long max_i = -1; for (i = 0; i < 5; i++) { if(occ_1_array[i] > max) { max = occ_1_array[i]; max_i = i; } } if(max_i == -1) { return 0; } if(max <= 1) { return 0; } ///if we have two max for (i = 0; i < 5; i++) { if(occ_1_array[i] == max && i != max_i) { return 0; } } long long new_0 = occ_0 + 1; long long new_total = sub_length + 1; ///note: here occ_0++ since the read itself has a type0 double available = new_0 + max; double threshold = 0.95; available = available/((double)(new_total)); if(available < threshold) { return 0; } ///if we just have one snp, we need to phase it carefully if(num_haplotype == 1) { ///we must have just 1 match and 1 mismatch ///any other types are not good if(new_0 + max != new_total) { return 0; } if(filter_snp(new_0, max, new_total) == 0) { return 0; } } /** if(filter_snp(new_0, max, new_total) == 0) { return 0; } **/ InsertSNPVector(hap, sub_list, sub_length, s_H[max_i]); /** fprintf(stderr, "new_0: %d, occ_0: %d, max: %d, max_i: %d, sub_length: %d, new_total: %d, available: %lf\n", new_0, occ_0, max, max_i, sub_length, new_total, available); for (i = 0; i < sub_length; i++) { fprintf(stderr, "i: %d, site: %d, type: %d, char: %c, ID: %d, name: %.*s\n", i, sub_list[i].site, sub_list[i].type, sub_list[i].misBase, sub_list[i].overlapID, Get_NAME_LENGTH((*R_INF), overlap_list->list[sub_list[i].overlapID].y_id), Get_NAME((*R_INF),overlap_list->list[sub_list[i].overlapID].y_id)); } fprintf(stderr, "\n"); **/ } int calculate_distance_snp_vector(int8_t *vector1, int8_t *vector2, int Len) { int i; for (i = 0; i < Len; i++) { if(vector1[i] != vector2[i]) { if ((vector1[i] == 0 || vector1[i] == 1) && (vector2[i] == 0 || vector2[i] == 1)) { return 1; } } } return 0; } void print_core_snp(haplotype_evdience_alloc* hap) { int i, j; for (i = 0; i < hap->core_snp; i++) { fprintf(stderr, "core(i): %d, site: %d, occ_0: %d, occ_1: %d, occ_2: %d, score: %d\n", i, hap->snp_stat[i].site, hap->snp_stat[i].occ_0, hap->snp_stat[i].occ_1, hap->snp_stat[i].occ_2, hap->snp_stat[i].score); int vectorID = hap->snp_stat[i].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); for (j = 0; j < hap->overlap; j++) { if(vector[j] == 0) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 1) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 2) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } } } void merge_snp_vectors(haplotype_evdience_alloc* hap, int diff_vector_ID) { int8_t *r_vector = Get_Result_SNP_Vector((*hap)); int vectorLen = Get_SNP_Vector_Length((*hap)); memset(r_vector, -1, vectorLen); hap->result_stat.occ_0 = 0; hap->result_stat.occ_1 = 0; int8_t* vector; int vectorID; int i, j; for (i = 0; i < hap->core_snp; i++) { if(i == diff_vector_ID) { continue; } vectorID = hap->snp_stat[i].id; vector = Get_SNP_Vector((*hap), vectorID); for (j = 0; j < vectorLen; j++) { if(r_vector[j] == -1) { if(vector[j] == 0) { hap->result_stat.occ_0++; r_vector[j] = vector[j]; } else if(vector[j] == 1) { hap->result_stat.occ_1++; r_vector[j] = vector[j]; } } else ///can debug here { if((vector[j] != -1 && vector[j] != 2 && vector[j] != r_vector[j])) { fprintf(stderr, "j: %d, vector[j]: %d, r_vector[j]: %d, hap->core_snp: %d, diff_vector_ID: %d\n", j, vector[j], r_vector[j], hap->core_snp, diff_vector_ID); print_core_snp(hap); } } } } hap->result_stat.overlap_num = hap->result_stat.occ_0 + hap->result_stat.occ_1; } void add_to_result_snp_vector(haplotype_evdience_alloc* hap, int8_t *new_vector, int Len) { int8_t *r_vector = Get_Result_SNP_Vector((*hap)); int j; for (j = 0; j < Len; j++) { if(r_vector[j] == -1) { if(new_vector[j] == 0) { hap->result_stat.occ_0++; r_vector[j] = new_vector[j]; } else if(new_vector[j] == 1) { hap->result_stat.occ_1++; r_vector[j] = new_vector[j]; } } ///can debug here } hap->result_stat.overlap_num = hap->result_stat.occ_0 + hap->result_stat.occ_1; } int debug_add_to_result_snp_vector(haplotype_evdience_alloc* hap, int8_t *new_vector, int Len) { int8_t *r_vector = Get_Result_SNP_Vector((*hap)); int j; for (j = 0; j < Len; j++) { if(r_vector[j] == -1) { if(new_vector[j] == 0) { hap->result_stat.occ_0++; r_vector[j] = new_vector[j]; } else if(new_vector[j] == 1) { hap->result_stat.occ_1++; r_vector[j] = new_vector[j]; } } else ///can debug here { if((new_vector[j] != -1 && new_vector[j] != 2 && new_vector[j] != r_vector[j])) { fprintf(stderr, "j: %d\n", j); return 0; } } } return 1; } int merge_snp_vectors_and_test(haplotype_evdience_alloc* hap, int diff_vector_ID) { int8_t *r_vector = Get_Result_SNP_Vector((*hap)); int vectorLen = Get_SNP_Vector_Length((*hap)); memset(r_vector, -1, vectorLen); hap->result_stat.occ_0 = 0; hap->result_stat.occ_1 = 0; int8_t* vector; int vectorID; int i, j; for (i = 0; i < hap->core_snp; i++) { if(i == diff_vector_ID) { continue; } vectorID = hap->snp_stat[i].id; vector = Get_SNP_Vector((*hap), vectorID); for (j = 0; j < vectorLen; j++) { if(r_vector[j] == -1) { if(vector[j] == 0) { hap->result_stat.occ_0++; r_vector[j] = vector[j]; } else if(vector[j] == 1) { hap->result_stat.occ_1++; r_vector[j] = vector[j]; } } else ///can debug here { ///has confilict if(vector[j] != -1 && vector[j] != 2 && vector[j] != r_vector[j]) { return 0; } } } } hap->result_stat.overlap_num = hap->result_stat.occ_0 + hap->result_stat.occ_1; return 1; } int generate_haplotypes(haplotype_evdience_alloc* hap) { int j; int vectorID, vectorID2; int diff_core_vector = 0; int diff_vector_ID = -1; int8_t *vector, *vector2; if(hap->core_snp == 0) { return 0; } qsort(hap->snp_stat, hap->available_snp, sizeof(SnpStats), cmp_snp_stats); ///the hap->core_snp is used to find centriod ///if there are <5 vectors in core_snp, we didn't allow different vector if (hap->core_snp < 5) { if(merge_snp_vectors_and_test(hap, -1) == 0) { return 0; } } else ///for vectors in core_snp, we allow at most one different vector when there are >= 5 vectors in core_snp { ///there are two condition: 1. vector 0 is the different one. 2. vector 0 is not the different one diff_vector_ID = -1; ///first try to merge all vector together if(merge_snp_vectors_and_test(hap, -1) == 0) { for (j = hap->core_snp - 1; j >= 0; j--) { if(merge_snp_vectors_and_test(hap, j) == 1) { diff_vector_ID = j; break; } } if(j == -1) { return 0; } } } ///after merge, we get result vector vector = Get_Result_SNP_Vector((*hap)); ///and for each non-core snp vector, if it has no conflict with result vector /// add it to result vector for (j = hap->core_snp; j < hap->available_snp; j++) { vectorID2 = hap->snp_stat[j].id; vector2 = Get_SNP_Vector((*hap), vectorID2); if(calculate_distance_snp_vector(vector, vector2, Get_SNP_Vector_Length((*hap))) == 0) { add_to_result_snp_vector(hap, vector2, Get_SNP_Vector_Length((*hap))); } } ///merge_snp_vectors(hap, diff_vector_ID); ///for read only have 1 snp, we need a more strict condition if (hap->core_snp == 1 && filter_one_snp(hap->result_stat.occ_0 + 1, hap->result_stat.occ_1, hap->result_stat.overlap_num + 1) == 0) { return 0; } return 1; } void print_snp_in_line(haplotype_evdience_alloc* hap) { int j, i; uint32_t* column; fprintf(stderr, "\n\n\n###########hap->available_snp: %d###########\n", hap->available_snp); for (j = 0; j < hap->available_snp; j++) { fprintf(stderr, "*********j: %d, site: %d, id: %d*********\n", j, hap->snp_stat[j].site, hap->snp_stat[j].id); fprintf(stderr, "type(0):\n"); int vectorID = hap->snp_stat[j].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); for (i = 0; i < hap->overlap; i++) { if(vector[i] == 0) { fprintf(stderr, "%3d, ", i); } } fprintf(stderr, "\n"); fprintf(stderr, "type(1):\n"); for (i = 0; i < hap->overlap; i++) { if(vector[i] == 1) { fprintf(stderr, "%3d, ", i); } } fprintf(stderr, "\n"); } fprintf(stderr, "***********************\n"); for (i = 0; i < hap->dp.snp_num; i++) { fprintf(stderr, "hap->dp.max[%d]: %d, hap->dp.backtrack_length: %d\n", i, hap->dp.max[i], hap->dp.backtrack_length[i]); if(hap->dp.backtrack_length[i] != 0) { column = Get_DP_Backtrack_Column(hap->dp, i); for (j = 0; j < hap->dp.backtrack_length[i]; j++) { fprintf(stderr, "pre: %d,", column[j]); } fprintf(stderr, "\n"); } } } int generate_haplotypes_DP(haplotype_evdience_alloc* hap, overlap_region_alloc* overlap_list, All_reads* R_INF) { int j, i; int vectorID, vectorID2; int diff_core_vector = 0; int diff_vector_ID = -1; int8_t *vector, *vector2; if(hap->available_snp == 0) { return 0; } /** fprintf(stderr, "*******\nhap->available_snp: %d\n", hap->available_snp); for (j = 0; j < hap->available_snp; j++) { fprintf(stderr, "site: %d, id: %d\n", hap->snp_stat[j].site, hap->snp_stat[j].id); } **/ ///if hap->available_snp == 1, the following codes would have bugs ///filter snps that are highly likly false if(hap->available_snp > 1) { i = 0; ///if a snp is very near to others, it should not be a real snp for (j = 0; j < hap->available_snp; j++) { if(j > 0 && j < hap->available_snp - 1) { if(hap->snp_stat[j].site != hap->snp_stat[j - 1].site + 1 && hap->snp_stat[j].site + 1 != hap->snp_stat[j + 1].site) { hap->snp_stat[i] = hap->snp_stat[j]; i++; } } else if(j == 0) { if(hap->snp_stat[j].site + 1 != hap->snp_stat[j + 1].site) { hap->snp_stat[i] = hap->snp_stat[j]; i++; } } else { if(hap->snp_stat[j].site != hap->snp_stat[j - 1].site + 1) { hap->snp_stat[i] = hap->snp_stat[j]; i++; } } } hap->available_snp = i; } int flag; long long overlap_length, total_read, unuseful_read; total_read = unuseful_read = 0; ///check if any read may be conflict with others for (i = 0; i < overlap_list->length; i++) { overlap_length = overlap_list->list[i].x_pos_e - overlap_list->list[i].x_pos_s + 1; if (overlap_length * OVERLAP_THRESHOLD <= overlap_list->list[i].align_length) { total_read++; flag = -1; for (j = 0; j < hap->available_snp; j++) { vectorID = hap->snp_stat[j].id; vector = Get_SNP_Vector((*hap), vectorID); if(vector[i] != 2 && flag == 2) { unuseful_read++; flag = 3; break; } if(vector[i] == 2) { flag = 2; } } if(flag == 3) { for (j = 0; j < hap->available_snp; j++) { vectorID = hap->snp_stat[j].id; vector = Get_SNP_Vector((*hap), vectorID); vector[i] = 2; } } } } /*******************************DP********************************/ init_DP_matrix(&(hap->dp), hap->available_snp); long long equal_best = 0; uint32_t* column; long long column_length; for (i = 0; i < hap->available_snp; i++) { ///vector of snp i vectorID = hap->snp_stat[i].id; vector = Get_SNP_Vector((*hap), vectorID); hap->dp.max[i] = 1; hap->dp.backtrack_length[i] = 0; equal_best = 0; column = Get_DP_Backtrack_Column(hap->dp, i); column_length = Get_DP_Backtrack_Column_Length(hap->dp, i); for (j = 0; j < i; j++) { ///vector of snp j vectorID2 = hap->snp_stat[j].id; vector2 = Get_SNP_Vector((*hap), vectorID2); ///vector is compatible with vector2 if(calculate_distance_snp_vector(vector, vector2, Get_SNP_Vector_Length((*hap))) == 0) { if(hap->dp.max[i] < hap->dp.max[j] + 1) { hap->dp.max[i] = hap->dp.max[j] + 1; column[0] = j; equal_best = 1; } else if(hap->dp.max[i] == hap->dp.max[j] + 1) { column[equal_best] = j; equal_best++; } } } hap->dp.backtrack_length[i] = equal_best; } /*******************************DP********************************/ /** vector = Get_Result_SNP_Vector((*hap)); memset(vector, -1, Get_SNP_Vector_Length((*hap))); vector = Get_Result_SNP_Vector((*hap)); for (i = 0; i < hap->available_snp; i++) { if(hap->dp.max[i] > 1) { vector = Get_Result_SNP_Vector((*hap)); memset(vector, -1, Get_SNP_Vector_Length((*hap))); if(hap->dp.backtrack_length[i] < 1) { fprintf(stderr, "error\n"); } int debug_i = i; int round = hap->dp.max[i]; while (hap->dp.max[debug_i] != 1) { vectorID2 = hap->snp_stat[debug_i].id; vector2 = Get_SNP_Vector((*hap), vectorID2); if(debug_add_to_result_snp_vector(hap, vector2, Get_SNP_Vector_Length((*hap))) == 0) { fprintf(stderr, "debug_i: %d\n", debug_i); print_snp_in_line(hap); } if(hap->dp.max[debug_i] != round) { fprintf(stderr, "hahah\n"); } debug_i = hap->dp.backtrack_length[0]; round--; } } } vector = Get_Result_SNP_Vector((*hap)); memset(vector, -1, Get_SNP_Vector_Length((*hap))); if(hap->available_snp > 4) { fprintf(stderr, "\n\n\n###########hap->available_snp: %d###########\n", hap->available_snp); for (j = 0; j < hap->available_snp; j++) { fprintf(stderr, "*********site: %d, id: %d*********\n", hap->snp_stat[j].site, hap->snp_stat[j].id); fprintf(stderr, "type(0):\n"); int vectorID = hap->snp_stat[j].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); for (i = 0; i < hap->overlap; i++) { if(vector[i] == 0) { fprintf(stderr, "%3d, ", i); } } fprintf(stderr, "\n"); fprintf(stderr, "type(1):\n"); for (i = 0; i < hap->overlap; i++) { if(vector[i] == 1) { fprintf(stderr, "%3d, ", i); } } fprintf(stderr, "\n"); } fprintf(stderr, "***********************\n"); for (i = 0; i < hap->dp.snp_num; i++) { fprintf(stderr, "hap->dp.max[%d]: %d, hap->dp.backtrack_length: %d\n", i, hap->dp.max[i], hap->dp.backtrack_length[i]); if(hap->dp.backtrack_length[i] != 0) { column = Get_DP_Backtrack_Column(hap->dp, i); for (j = 0; j < hap->dp.backtrack_length[i]; j++) { fprintf(stderr, "pre: %d,", column[j]); } fprintf(stderr, "\n"); } } } **/ /** fprintf(stderr, "overlap_list->length: %u, total_read: %u, unuseful_read: %u\n", overlap_list->length, total_read, unuseful_read); for (i = 0; i < overlap_list->length; i++) { overlap_length = overlap_list->list[i].x_pos_e - overlap_list->list[i].x_pos_s + 1; if (overlap_length * OVERLAP_THRESHOLD <= overlap_list->list[i].align_length) { total_read++; flag = -1; for (j = 0; j < hap->available_snp; j++) { vectorID = hap->snp_stat[j].id; vector = Get_SNP_Vector((*hap), vectorID); if(vector[i] != 2 && flag == 2) { fprintf(stderr, "hahahah\n"); break; } if(vector[i] == 2) { flag = 2; } } } } **/ return 0; } void print_Haplotype(haplotype_evdience_alloc* hap, overlap_region_alloc* overlap_list, All_reads* R_INF) { int j, i; fprintf(stderr, "\nhap->snp: %d, hap->length: %d, perc: %d, x_name: %.*s\n", hap->snp, hap->length, (hap->snp == 0? 0: hap->length/hap->snp), Get_NAME_LENGTH((*R_INF), overlap_list->list[0].x_id), Get_NAME((*R_INF),overlap_list->list[0].x_id) ); fprintf(stderr, "hap->available_snp: %d, hap->core_snp:%d\n", hap->available_snp, hap->core_snp); int Len_x, matched_overlap = 0; for (j = 0; j < overlap_list->length; j++) { Len_x = overlap_list->list[j].x_pos_e - overlap_list->list[j].x_pos_s + 1; if (Len_x * OVERLAP_THRESHOLD <= overlap_list->list[j].align_length) { matched_overlap++; } } fprintf(stderr, "occ_0: %d, occ_1: %d, overlap_num: %d, matched_overlap: %d, overlap_list->length: %d\n", hap->result_stat.occ_0, hap->result_stat.occ_1, hap->result_stat.overlap_num, matched_overlap, overlap_list->length); return; fprintf(stderr, "Phaseing sucessfully!\n"); fprintf(stderr, "occ_0: %d, occ_1: %d, overlap_num: %d\n", hap->result_stat.occ_0, hap->result_stat.occ_1, hap->result_stat.overlap_num); int8_t* vector = Get_Result_SNP_Vector((*hap)); for (j = 0; j < hap->overlap; j++) { if(vector[j] == 0) { fprintf(stderr, "Ptype: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 1) { fprintf(stderr, "Ptype: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 2) { fprintf(stderr, "Ptype: %d, ID: %d\n", vector[j], j); } } for (i = 0; i < hap->core_snp; i++) { fprintf(stderr, "core(i): %d, site: %d, occ_0: %d, occ_1: %d, occ_2: %d, score: %d\n", i, hap->snp_stat[i].site, hap->snp_stat[i].occ_0, hap->snp_stat[i].occ_1, hap->snp_stat[i].occ_2, hap->snp_stat[i].score); int vectorID = hap->snp_stat[i].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); for (j = 0; j < hap->overlap; j++) { if(vector[j] == 0) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 1) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 2) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } } for (; i < hap->available_snp; i++) { fprintf(stderr, "i: %d, site: %d, occ_0: %d, occ_1: %d, occ_2: %d, score: %d\n", i, hap->snp_stat[i].site, hap->snp_stat[i].occ_0, hap->snp_stat[i].occ_1, hap->snp_stat[i].occ_2, hap->snp_stat[i].score); int vectorID = hap->snp_stat[i].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); for (j = 0; j < hap->overlap; j++) { if(vector[j] == 0) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 1) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } for (j = 0; j < hap->overlap; j++) { if(vector[j] == 2) { fprintf(stderr, "type: %d, ID: %d\n", vector[j], j); } } } } void debug_near_snp(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, Correct_dumy* dumy, haplotype_evdience_alloc* hap) { int i, j, overlap_length, window_start, window_end, flag; long long window_num = (g_read->length + WINDOW - 1) / WINDOW; int total_read = 0; int unuseful_read = 0; for (i = 0; i < overlap_list->length; i++) { int flag = -1; overlap_length = overlap_list->list[i].x_pos_e - overlap_list->list[i].x_pos_s + 1; if (overlap_length * OVERLAP_THRESHOLD <= overlap_list->list[i].align_length) { total_read++; for (j = 0; j < hap->available_snp; j++) { int vectorID = hap->snp_stat[j].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); if(vector[i] != 2 && flag == 2) { unuseful_read++; break; } if(vector[i] == 2) { flag = 2; } } } } for (j = 1; j < hap->available_snp; j++) { if(hap->snp_stat[j].site <= hap->snp_stat[j - 1].site) { fprintf(stderr, "error\n"); } } for (j = 0; j < hap->available_snp; j++) { fprintf(stderr, "\n\nsite: %d, score: %d\n", hap->snp_stat[j].site, hap->snp_stat[j].score ); if((j>0 && hap->snp_stat[j].site == hap->snp_stat[j - 1].site + 1) || (j < hap->available_snp - 1 && hap->snp_stat[j].site + 1 == hap->snp_stat[j + 1].site)) { fprintf(stderr, "x_name: %.*s\n", Get_NAME_LENGTH((*R_INF), overlap_list->list[0].x_id), Get_NAME((*R_INF),overlap_list->list[0].x_id)); int vectorID = hap->snp_stat[j].id; int8_t* vector = Get_SNP_Vector((*hap), vectorID); for (i = 0; i < hap->overlap; i++) { if(vector[i] == 0) { fprintf(stderr, "type: %d, ID: %d\n", vector[i], i); } } for (i = 0; i < hap->overlap; i++) { if(vector[i] == 1) { fprintf(stderr, "type: %d, ID: %d, %.*s\n", vector[i], i, Get_NAME_LENGTH((*R_INF), overlap_list->list[i].y_id), Get_NAME((*R_INF),overlap_list->list[i].y_id)); } } window_start = 0; window_end = WINDOW - 1; if (window_end >= g_read->length) { window_end = g_read->length - 1; } long long ijk; for (ijk = 0; ijk < window_num; ijk++) { if(hap->snp_stat[j].site <= window_end && hap->snp_stat[j].site >= window_start) { fprintf(stderr, "winID: %d, winNum: %d, window_start: %d, window_end: %d\n", ijk, window_num, window_start, window_end); dumy->length = 0; dumy->lengthNT = 0; ///flag返回的是重叠数量 ///dumy->length返回的是有效完全重叠的数量 ///dumy->lengthNT返回的是有效不完全重叠的数量 ///return overlaps that is overlaped with [window_start, window_end] flag = get_available_interval(window_start, window_end, overlap_list, dumy); switch (flag) { case 1: ///找到匹配 break; case 0: ///没找到匹配 break; case -2: ///下一个window也不会存在匹配, 直接跳出 i = window_num; break; } for (i = 0; i < dumy->length; i++) { ///这个是那个overlap的ID,而不是overlap里对应窗口的ID int overlapID = dumy->overlapID[i]; ///overlap_list->list[overlapID].x_pos_s is the begining of the whole overlap int correct_x_pos_s = (overlap_list->list[overlapID].x_pos_s / WINDOW) * WINDOW; ///window_start is the begining of this window in the whole x_read int windowID = (window_start - correct_x_pos_s) / WINDOW; ///如果这个window不匹配,跳过 if (overlap_list->list[overlapID].w_list[windowID].y_end == -1) { continue; } if(vector[overlapID] == 1) { fprintf(stderr, "overlapID: %d, x_strat: %d, x_end: %d, y_start: %d, y_end: %d\n", overlapID, overlap_list->list[overlapID].w_list[windowID].x_start, overlap_list->list[overlapID].w_list[windowID].x_end, overlap_list->list[overlapID].w_list[windowID].y_start, overlap_list->list[overlapID].w_list[windowID].y_end); recover_UC_Read_sub_region(dumy->overlap_region, overlap_list->list[overlapID].w_list[windowID].y_start, overlap_list->list[overlapID].w_list[windowID].y_end -overlap_list->list[overlapID].w_list[windowID].y_start + 1, overlap_list->list[overlapID].y_pos_strand, R_INF, overlap_list->list[overlapID].y_id); char* x_string = g_read->seq + overlap_list->list[overlapID].w_list[windowID].x_start; char* y_string = dumy->overlap_region; fprintf(stderr, "x_string: \n%.*s\n", overlap_list->list[overlapID].w_list[windowID].x_end - overlap_list->list[overlapID].w_list[windowID].x_start + 1, x_string); fprintf(stderr, "y_string: \n%.*s\n", overlap_list->list[overlapID].w_list[windowID].y_end - overlap_list->list[overlapID].w_list[windowID].y_start + 1, y_string); int haha_i = 0; for (haha_i = 0; haha_i < overlap_list->list[overlapID].w_list[windowID].cigar.length; haha_i++) { fprintf(stderr, "oper: %d, len: %d\n", overlap_list->list[overlapID].w_list[windowID].cigar.C_C[haha_i], overlap_list->list[overlapID].w_list[windowID].cigar.C_L[haha_i] ); } } } } window_start = window_start + WINDOW; window_end = window_end + WINDOW; if (window_end >= g_read->length) { window_end = g_read->length - 1; } } } } fprintf(stderr, "total_read: %d, unuseful_read: %d\n", total_read, unuseful_read); } void partition_overlaps(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, Correct_dumy* dumy, haplotype_evdience_alloc* hap) { ResizeInitHaplotypeEvdience(hap); long long window_num = (g_read->length + WINDOW - 1) / WINDOW; long long i, j, overlap_length; long long window_start, window_end; long long num_availiable_win = 0; window_start = 0; window_end = WINDOW - 1; if (window_end >= g_read->length) { window_end = g_read->length - 1; } int flag; for (i = 0; i < window_num; i++) { dumy->length = 0; dumy->lengthNT = 0; ///flag返回的是重叠数量 ///dumy->length返回的是有效完全重叠的数量 ///dumy->lengthNT返回的是有效不完全重叠的数量 ///return overlaps that is overlaped with [window_start, window_end] flag = get_available_interval(window_start, window_end, overlap_list, dumy); switch (flag) { case 1: ///找到匹配 break; case 0: ///没找到匹配 break; case -2: ///下一个window也不会存在匹配, 直接跳出 i = window_num; break; } ///这个是available overlap里所有window的数量... ///num_availiable_win = num_availiable_win + dumy->length + dumy->lengthNT; num_availiable_win = num_availiable_win + dumy->length; cluster(g_read->seq, window_start, window_end, overlap_list, dumy, R_INF, hap); window_start = window_start + WINDOW; window_end = window_end + WINDOW; if (window_end >= g_read->length) { window_end = g_read->length - 1; } } ///very time-consuming ///it seems that hap->list has alread been sorted by site qsort(hap->list, hap->length, sizeof(haplotype_evdience), cmp_haplotype_evdience); ///debug_hap_information(overlap_list, R_INF, g_read, hap, dumy); SetSnpMatrix(hap, hap->snp, overlap_list->length); uint64_t pre_site = (uint64_t)-1; uint64_t num_of_snps = 0; long long pre_i = -1; long long sub_length; haplotype_evdience* sub_list; ///long long debug_total_length = 0; ////split reads for (i = 0; i < hap->length; i++) { if(pre_site != hap->list[i].site) { if(i != 0) { sub_list = hap->list + pre_i; sub_length = i - pre_i; ///debug_total_length = debug_total_length + sub_length; split_sub_list(hap, sub_list, sub_length, hap->snp, overlap_list, R_INF); } num_of_snps++; pre_site = hap->list[i].site; pre_i = i; } } if(pre_i != -1) { sub_list = hap->list + pre_i; sub_length = i - pre_i; ///debug_total_length = debug_total_length + sub_length; split_sub_list(hap, sub_list, sub_length, hap->snp, overlap_list, R_INF); } ///debug_snp_matrix(hap); generate_haplotypes_DP(hap, overlap_list, R_INF); ///debug_near_snp(overlap_list, R_INF, g_read, dumy, hap); ///debug_snp_matrix(hap); if(generate_haplotypes(hap)) { //print_Haplotype(hap, overlap_list, R_INF); ///if we can phase, we need to exclude the reads of different haplotype int8_t* vector = Get_Result_SNP_Vector((*hap)); for (j = 0; j < Get_SNP_Vector_Length((*hap)); j++) { if(vector[j] == 1) { overlap_list->list[j].align_length = 0; } } } } void correct_overlap(overlap_region_alloc* overlap_list, All_reads* R_INF, UC_Read* g_read, Correct_dumy* dumy, UC_Read* overlap_read, Graph* g, long long* matched_overlap_0, long long* matched_overlap_1, long long* potiental_matched_overlap_0, long long* potiental_matched_overlap_1, Cigar_record* current_cigar, haplotype_evdience_alloc* hap, Round2_alignment* second_round) { reverse_complement(g_read->seq, g_read->length); clear_Correct_dumy(dumy, overlap_list); long long window_num = (g_read->length + WINDOW - 1) / WINDOW; long long i; long long window_start, window_end; window_start = 0; window_end = WINDOW - 1; if (window_end >= g_read->length) { window_end = g_read->length - 1; } int flag; for (i = 0; i < window_num; i++) { dumy->length = 0; dumy->lengthNT = 0; flag = get_interval(window_start, window_end, overlap_list, dumy); switch (flag) { case 1: ///找到匹配 break; case 0: ///没找到匹配 break; case -2: ///下一个window也不会存在匹配, 直接跳出 i = window_num; break; } if(dumy->length + dumy->lengthNT>overlap_list->length) { fprintf(stderr, "error length\n"); } ///verify_get_interval(window_start, window_end, overlap_list, dumy); verify_window(window_start, window_end, overlap_list, dumy, R_INF, g_read->seq); window_start = window_start + WINDOW; window_end = window_end + WINDOW; if (window_end >= g_read->length) { window_end = g_read->length - 1; } } debug_stats(overlap_list, R_INF, g_read, dumy, overlap_read, potiental_matched_overlap_0, potiental_matched_overlap_1); recalcate_window(overlap_list, R_INF, g_read, dumy, overlap_read); debug_stats(overlap_list, R_INF, g_read, dumy, overlap_read, matched_overlap_0, matched_overlap_1); partition_overlaps(overlap_list, R_INF, g_read, dumy, hap); generate_consensus(overlap_list, R_INF, g_read, dumy, g, current_cigar, second_round); /** ///fprintf(stderr, "length: %lld, corrected_base: %lld\n", g_read->length, dumy->corrected_base); EdlibAlignResult result = edlibAlign(g_read->seq, g_read->length, dumy->corrected_read, dumy->corrected_read_length, edlibNewAlignConfig(-1, EDLIB_MODE_NW, EDLIB_TASK_PATH, NULL, 0)); if (result.status == EDLIB_STATUS_OK) { if (dumy->corrected_base == 0 && result.editDistance!= 0) { fprintf(stderr, "error 0\n"); } if (dumy->corrected_base < result.editDistance) { fprintf(stderr, "error 1\n"); } // fprintf(stderr, "****\n distance: %d, alignmentLength: %d, startLocations: %d, endLocations: %d, corrected_base: %d\n", // result.editDistance, result.alignmentLength, result.startLocations[0], result.endLocations[0], dumy->corrected_base); // char* cigar = edlibAlignmentToCigar(result.alignment, result.alignmentLength, EDLIB_CIGAR_STANDARD); // fprintf(stderr,"%s\n", cigar); // free(cigar); } else { fprintf(stderr, "error\n"); } edlibFreeAlignResult(result); for (i = 0; i < dumy->corrected_read_length; i++) { if (dumy->corrected_read[i] != 'A' && dumy->corrected_read[i] != 'C' && dumy->corrected_read[i] != 'G' && dumy->corrected_read[i] != 'T') { fprintf(stderr, "error\n"); } } **/ } void init_Cigar_record(Cigar_record* dummy) { dummy->length = 0; dummy->size = 100; dummy->record = (uint32_t*)malloc(sizeof(uint32_t)*dummy->size); dummy->lost_base_length = 0; dummy->lost_base_size = 100; dummy->lost_base = (char*)malloc(sizeof(char)*dummy->lost_base_size); dummy->current_operation_length = 0; dummy->current_operation = 127; } void destory_Cigar_record(Cigar_record* dummy) { free(dummy->record); free(dummy->lost_base); } void clear_Cigar_record(Cigar_record* dummy) { dummy->new_read_length = 0; dummy->length = 0; dummy->lost_base_length = 0; dummy->current_operation_length = 0; dummy->current_operation = 127; } void init_Correct_dumy(Correct_dumy* list) { list->size = 0; list->length = 0; list->lengthNT = 0; list->start_i = 0; list->overlapID = NULL; int i; for (i = 0; i < 256; i++) { list->Peq_SSE[i] = _mm_setzero_si128(); } list->corrected_read_size = 1000; list->corrected_read_length = 0; list->corrected_read = (char*)malloc(sizeof(char)*list->corrected_read_size); list->corrected_base = 0; } void destory_Correct_dumy(Correct_dumy* list) { free(list->overlapID); free(list->corrected_read); } void clear_Correct_dumy(Correct_dumy* list, overlap_region_alloc* overlap_list) { list->length = 0; list->lengthNT = 0; list->start_i = 0; if (list->size < overlap_list->length) { list->size = overlap_list->length; list->overlapID = (uint64_t*)realloc(list->overlapID, list->size*sizeof(uint64_t)); } list->last_boundary_length = 0; list->corrected_read_length = 0; list->corrected_base = 0; } void clear_Correct_dumy_pure(Correct_dumy* list) { list->length = 0; list->lengthNT = 0; list->start_i = 0; list->last_boundary_length = 0; list->corrected_read_length = 0; list->corrected_base = 0; } void pre_filter_by_nearby_single(k_mer_pos* new_n_list, k_mer_pos* old_n_list, uint64_t n_length, uint64_t n_end_pos, UC_Read* g_read, All_reads* R_INF, Correct_dumy* dumy, uint64_t* new_n_length) { (*new_n_length) = 0; ///这种就是0,new_n_list也不需要有数据 if (n_length == 0) { return; } char* x_string = NULL; char* y_string = NULL; long long x_offset = (long long)(n_end_pos) - WINDOW + 1; if (x_offset < 0) { x_offset = 0; } ///x_length是x上待验证区间的实际长度 ///x_offset是x上待验证区间的起始位置 ///如果是向前取待验证区间,那么x_length至少大于等于40 (compressed k-mer长度,不是实际长度) long long x_length = n_end_pos - x_offset + 1; ///这种也不要过滤了,直接把old_n_list全部赋值过来就好了 if (x_length < (WINDOW/2)) { (*new_n_length) = n_length; memcpy(new_n_list, old_n_list, sizeof(k_mer_pos)*n_length); return; } long long Window_Len = x_length + (THRESHOLD << 1); x_string = g_read->seq + x_offset; long long y_offset; long long y_length; long long i = 0; long long y_read_length; int end_site; unsigned int error; for (i = 0; i < n_length; i++) { ////old_n_list[i].offset是y上k-mer的结束位置 ///n_end_pos是x上k-mer的结束位置 ///x_length是x上区间长度 y_offset = (long long)(old_n_list[i].offset) - x_length; ///这种情况下弃疗 if (y_offset < 0) { new_n_list[(*new_n_length)].readID = old_n_list[i].readID; new_n_list[(*new_n_length)].offset = old_n_list[i].offset; (*new_n_length)++; continue; } y_offset = y_offset - THRESHOLD; ///还能抢救 if (y_offset < 0) { y_offset = 0; } ///y的read的总长度 y_read_length = Get_READ_LENGTH((*R_INF), old_n_list[i].readID); y_length = MIN(Window_Len, y_read_length - y_offset); ///如果y的长度比x长度还小,那就直接弃疗了 if (y_length < x_length) { new_n_list[(*new_n_length)].readID = old_n_list[i].readID; new_n_list[(*new_n_length)].offset = old_n_list[i].offset; (*new_n_length)++; continue; } ///y的方向都是0,因为索引里都是0 recover_UC_Read_sub_region(dumy->overlap_region, y_offset, y_length, 0, R_INF, old_n_list[i].readID); y_string = dumy->overlap_region; memset (y_string + y_length, 0, Window_Len - y_length); end_site = Reserve_Banded_BPM(y_string, y_length, x_string, x_length, THRESHOLD, &error); if (error!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = old_n_list[i].readID; new_n_list[(*new_n_length)].offset = old_n_list[i].offset; (*new_n_length)++; } } } void pre_filter_by_nearby(k_mer_pos* new_n_list, k_mer_pos* old_n_list, uint64_t n_length, uint64_t n_end_pos, UC_Read* g_read, All_reads* R_INF, Correct_dumy* dumy, uint64_t* new_n_length) { (*new_n_length) = 0; ///这种就是0,new_n_list也不需要有数据 if (n_length == 0) { return; } char* x_string = NULL; char* y_string = NULL; long long x_offset = (long long)(n_end_pos) - WINDOW + 1; if (x_offset < 0) { x_offset = 0; } ///x_length是x上待验证区间的实际长度 ///x_offset是x上待验证区间的起始位置 ///如果是向前取待验证区间,那么x_length至少大于等于40 (compressed k-mer长度,不是实际长度) long long x_length = n_end_pos - x_offset + 1; ///这种也不要过滤了,直接把old_n_list全部赋值过来就好了 if (x_length < (WINDOW/2)) { (*new_n_length) = n_length; memcpy(new_n_list, old_n_list, sizeof(k_mer_pos)*n_length); return; } long long Window_Len = x_length + (THRESHOLD << 1); x_string = g_read->seq + x_offset; long long y_offset; long long y_length; long long i = 0; long long y_read_length; int end_site; unsigned int error; int groupLen = 0; int return_sites[GROUP_SIZE]; unsigned int return_sites_error[GROUP_SIZE]; uint64_t readID[GROUP_SIZE]; uint64_t offset[GROUP_SIZE]; for (i = 0; i < n_length; i++) { ////old_n_list[i].offset是y上k-mer的结束位置 ///n_end_pos是x上k-mer的结束位置 ///x_length是x上区间长度 y_offset = (long long)(old_n_list[i].offset) - x_length; ///这种情况下弃疗 if (y_offset < 0) { new_n_list[(*new_n_length)].readID = old_n_list[i].readID; new_n_list[(*new_n_length)].offset = old_n_list[i].offset; (*new_n_length)++; continue; } y_offset = y_offset - THRESHOLD; ///还能抢救 if (y_offset < 0) { y_offset = 0; } ///y的read的总长度 y_read_length = Get_READ_LENGTH((*R_INF), old_n_list[i].readID); y_length = MIN(Window_Len, y_read_length - y_offset); ///如果y的长度比x长度还小,那就直接弃疗了 if (y_length < x_length) { new_n_list[(*new_n_length)].readID = old_n_list[i].readID; new_n_list[(*new_n_length)].offset = old_n_list[i].offset; (*new_n_length)++; continue; } if(y_length == Window_Len) { ///y的方向都是0,因为索引里都是0 recover_UC_Read_sub_region(dumy->overlap_region_group[groupLen], y_offset, y_length, 0, R_INF, old_n_list[i].readID); readID[groupLen] = old_n_list[i].readID; offset[groupLen] = old_n_list[i].offset; groupLen++; if (groupLen == GROUP_SIZE) { Reserve_Banded_BPM_4_SSE_only(dumy->overlap_region_group[0], dumy->overlap_region_group[1], dumy->overlap_region_group[2], dumy->overlap_region_group[3], y_length, x_string, x_length, return_sites, return_sites_error, THRESHOLD, dumy->Peq_SSE); groupLen = 0; if (return_sites_error[0]!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = readID[0]; new_n_list[(*new_n_length)].offset = offset[0]; (*new_n_length)++; } if (return_sites_error[1]!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = readID[1]; new_n_list[(*new_n_length)].offset = offset[1]; (*new_n_length)++; } if (return_sites_error[2]!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = readID[2]; new_n_list[(*new_n_length)].offset = offset[2]; (*new_n_length)++; } if (return_sites_error[3]!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = readID[3]; new_n_list[(*new_n_length)].offset = offset[3]; (*new_n_length)++; } } } else { ///y的方向都是0,因为索引里都是0 recover_UC_Read_sub_region(dumy->overlap_region, y_offset, y_length, 0, R_INF, old_n_list[i].readID); y_string = dumy->overlap_region; memset (y_string + y_length, 0, Window_Len - y_length); end_site = Reserve_Banded_BPM(y_string, y_length, x_string, x_length, THRESHOLD, &error); if (error!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = old_n_list[i].readID; new_n_list[(*new_n_length)].offset = old_n_list[i].offset; (*new_n_length)++; } } } if (groupLen == 1) { end_site = Reserve_Banded_BPM(dumy->overlap_region_group[0], Window_Len, x_string, x_length, THRESHOLD, &error); if (error!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = readID[0]; new_n_list[(*new_n_length)].offset = offset[0]; (*new_n_length)++; } } else { Reserve_Banded_BPM_4_SSE_only(dumy->overlap_region_group[0], dumy->overlap_region_group[1], dumy->overlap_region_group[2], dumy->overlap_region_group[3], Window_Len, x_string, x_length, return_sites, return_sites_error, THRESHOLD, dumy->Peq_SSE); for (i = 0; i < groupLen; i++) { if (return_sites_error[i]!=(unsigned int)-1) { new_n_list[(*new_n_length)].readID = readID[i]; new_n_list[(*new_n_length)].offset = offset[i]; (*new_n_length)++; } } groupLen = 0; } ///要排序.... if ((*new_n_length)>1) { qsort(new_n_list, (*new_n_length), sizeof(k_mer_pos), cmp_k_mer_pos); } } /**********************for prefilter************************ */ void destory_k_mer_pos_list_alloc_prefilter(k_mer_pos_list_alloc* list) { long long i = 0; for (i = 0; i < list->size; i++) { if (list->list[i].size != 0) { free(list->list[i].list); } } free(list->list); } void append_k_mer_pos_list_alloc_prefilter(k_mer_pos_list_alloc* list, k_mer_pos* n_list, uint64_t n_length, uint64_t n_end_pos, uint8_t n_direction, UC_Read* g_read, All_reads* R_INF, Correct_dumy* dumy) { if (list->length + 1 > list->size) { list->size = list->size * 2; list->list = (k_mer_pos_list*)realloc(list->list, sizeof(k_mer_pos_list)*list->size); ///新分配空间要初始化 memset(list->list + (list->size/2), 0, sizeof(k_mer_pos_list)*(list->size/2)); } if (list->list[list->length].size < n_length) { list->list[list->length].size = n_length; list->list[list->length].list = (k_mer_pos*)realloc(list->list[list->length].list, sizeof(k_mer_pos)*list->list[list->length].size); } ///list->list[list->length].list = n_list; ///memcpy(list->list[list->length].list, n_list, sizeof(k_mer_pos)*n_length); pre_filter_by_nearby(list->list[list->length].list, n_list, n_length, n_end_pos, g_read, R_INF, dumy, &n_length); if(n_length > 0) { list->list[list->length].length = n_length; list->list[list->length].direction = n_direction; list->list[list->length].end_pos = n_end_pos; list->length++; } } /**********************for prefilter************************ */