Release minimap2-2.6 (r623)

* Don't preallocate sdust_buf or minizer list. kalloc should be fast enough -
  benchmarks needed to confirm.

* Fixed a memory leak caused by divergence estimate (post v2.5)

* Reset the kalloc buffer after mapping a long query. This reduces peak memory
  when large chunks of memory are allocated, at the cost of performance, though.

Makefile

@@ -1,7 +1,7 @@
 CFLAGS=		-g -Wall -O2 -Wc++-compat
 CPPFLAGS=	-DHAVE_KALLOC
 INCLUDES=
-OBJS=		kthread.o kalloc.o misc.o bseq.o sketch.o sdust.o index.o chain.o align.o hit.o map.o format.o pe.o ksw2_ll_sse.o
+OBJS=		kthread.o kalloc.o misc.o bseq.o sketch.o sdust.o index.o chain.o align.o hit.o map.o format.o pe.o esterr.o ksw2_ll_sse.o
 PROG=		minimap2
 PROG_EXTRA=	sdust minimap2-lite
 LIBS=		-lm -lz -lpthread

NEWS.md

@@ -1,3 +1,58 @@
+Release 2.6-r623 (12 December 2017)
+-----------------------------------
+
+This release adds several features and fixes two minor bugs:
+
+ * Optionally build an index without sequences. This helps to reduce the
+   peak memory for read overlapping and is automatically applied when
+   base-level alignment is not requested.
+
+ * Approximately estimate per-base sequence divergence (i.e. 1-identity)
+   without performing base-level alignment, using a MashMap-like method. The
+   estimate is written to a new dv:f tag.
+
+ * Reduced the number of tiny terminal exons in RNA-seq alignment. The current
+   setting is conservative. Increase --end-seed-pen to drop more such exons.
+
+ * Reduced the peak memory when aligning long query sequences.
+
+ * Fixed a bug that is caused by HPC minimizers longer than 256bp. This should
+   have no effect in practice, but it is recommended to rebuild HPC indices if
+   possible.
+
+ * Fixed a bug when identifying identical hits (#71). This should only affect
+   artifactual reference consisting of near identical sequences.
+
+For genomic sequences, minimap2 should give nearly identical alignments to
+v2.5, except the new dv:f tag.
+
+(2.6: 12 December 2017, r623)
+
+
+
+Release 2.5-r572 (11 November 2017)
+-----------------------------------
+
+This release fixes several bugs and brings a couple of minor improvements:
+
+ * Fixed a severe bug that leads to incorrect mapping coordinates in rare
+   corner cases.
+
+ * Fixed underestimated mapping quality for chimeric alignments when the whole
+   query sequence contain many repetitive minimizers, and for chimeric
+   alignments caused by Z-drop.
+
+ * Fixed two bugs in Python binding: incorrect strand field (#57) and incorrect
+   sequence names for Python3 (#55).
+
+ * Improved mapping accuracy for highly overlapping paired ends.
+
+ * Added option -Y to use soft clipping for supplementary alignments (#56).
+
+(2.5: 11 November 2017, r572)
+
+
+
 Release 2.4-r555 (6 November 2017)
 ----------------------------------
 

README.md

@@ -1,10 +1,7 @@
-[![Release](https://img.shields.io/badge/Release-v2.4-blue.svg?style=flat)](https://github.com/lh3/minimap2/releases)
+[![GitHub Downloads](https://img.shields.io/github/downloads/lh3/minimap2/total.svg?style=social&logo=github&label=Download)](https://github.com/lh3/minimap2/releases)
-[![BioConda](https://img.shields.io/conda/vn/bioconda/minimap2.svg?style=flat)](https://anaconda.org/bioconda/minimap2)
+[![BioConda Install](https://img.shields.io/conda/dn/bioconda/minimap2.svg?style=flag&label=BioConda%20install)](https://anaconda.org/bioconda/minimap2)
 [![PyPI](https://img.shields.io/pypi/v/mappy.svg?style=flat)](https://pypi.python.org/pypi/mappy)
-[![Python Version](https://img.shields.io/pypi/pyversions/mappy.svg?style=flat)](https://pypi.python.org/pypi/mappy)
-[![License](https://img.shields.io/badge/License-MIT-blue.svg?style=flat)](LICENSE.txt)
 [![Build Status](https://travis-ci.org/lh3/minimap2.svg?branch=master)](https://travis-ci.org/lh3/minimap2)
-[![Downloads](https://img.shields.io/github/downloads/lh3/minimap2/total.svg?style=flat)](https://github.com/lh3/minimap2/releases)
 ## <a name="started"></a>Getting Started
 ```sh
 git clone https://github.com/lh3/minimap2
@@ -22,7 +19,7 @@ cd minimap2 && make
 ./minimap2 -ax splice -k14 -uf ref.fa reads.fa > aln.sam  # Nanopore Direct RNA-seq
 ./minimap2 -cx asm5 asm1.fa asm2.fa > aln.paf             # intra-species asm-to-asm alignment
 ./minimap2 -x ava-pb reads.fa reads.fa > overlaps.paf     # PacBio read overlap
-./minimap2 -x ava-one reads.fa reads.fa > overlaps.paf    # Nanopore read overlap
+./minimap2 -x ava-ont reads.fa reads.fa > overlaps.paf    # Nanopore read overlap
 # man page for detailed command line options
 man ./minimap2.1
 ```
@@ -71,9 +68,9 @@ Detailed evaluations are available from the [minimap2 preprint][preprint].
 Minimap2 only works on x86-64 CPUs. You can acquire precompiled binaries from
 the [release page][release] with:
 ```sh
-wget --no-check-certificate -O- https://github.com/lh3/minimap2/releases/download/v2.4/minimap2-2.4_x64-linux.tar.bz2 \
+curl -L https://github.com/lh3/minimap2/releases/download/v2.6/minimap2-2.6_x64-linux.tar.bz2 \
   | tar -jxvf -
-./minimap2-2.4_x64-linux/minimap2
+./minimap2-2.6_x64-linux/minimap2
 ```
 If you want to compile from the source, you need to have a C compiler, GNU make
 and zlib development files installed. Then type `make` in the source code
@@ -154,6 +151,20 @@ slight improvement to accuracy. For noisy Nanopore Direct RNA-seq reads, it is
 recommended to use a smaller k-mer size for increased sensitivity to the first
 or the last exons.
 
+Minimap2 rates an alignment by the score of the max-scoring sub-segment,
+*excluding* introns, and marks the best alignment as primary in SAM. When a
+spliced gene also has unspliced pseudogenes, minimap2 does not intentionally
+prefer spliced alignment, though in practice it more often marks the spliced
+alignment as the primary. By default, minimap2 outputs up to five secondary
+alignments (i.e. likely pseudogenes in the context of RNA-seq mapping). This
+can be tuned with option **-N**.
+
+For long RNA-seq reads, minimap2 may produce chimeric alignments potentially
+caused by gene fusions/structural variations or by an intron longer than the
+max intron length **-G** (200k by default). For now, it is not recommended to
+apply an excessively large **-G** as this slows down minimap2 and sometimes
+leads to false alignments.
+
 It is worth noting that by default `-x splice` prefers GT[A/G]..[C/T]AG
 over GT[C/T]..[A/G]AG, and then over other splicing signals. Considering
 one additional base improves the junction accuracy for noisy reads, but
@@ -309,7 +320,7 @@ highlighted in bold. The description may help to tune minimap2 parameters.
 
 ### <a name="help"></a>Getting help
 
-Manpage [minimap2.1](minimap2.1) provides detailed description of minimap2
+Manpage [minimap2.1][manpage] provides detailed description of minimap2
 command line options and optional tags. If you encounter bugs or have further
 questions or requests, you can raise an issue at the [issue page][issue].
 There is not a specific mailing list for the time being.
@@ -363,3 +374,4 @@ warmly welcomed.
 [mappyconda]: https://anaconda.org/bioconda/mappy
 [issue]: https://github.com/lh3/minimap2/issues
 [k8]: https://github.com/attractivechaos/k8
+[manpage]: https://lh3.github.io/minimap2/minimap2.html

align.c

@@ -1,6 +1,7 @@
 #include <assert.h>
 #include <string.h>
 #include <stdlib.h>
+#include <math.h>
 #include "minimap.h"
 #include "mmpriv.h"
 #include "ksw2.h"
@@ -110,14 +111,17 @@ static void mm_fix_cigar(mm_reg1_t *r, const uint8_t *qseq, const uint8_t *tseq,
 	}
 	if ((p->cigar[0]&0xf) == 1 || (p->cigar[0]&0xf) == 2) { // get rid of leading I or D
 		int32_t l = p->cigar[0] >> 4;
-		if ((p->cigar[0]&0xf) == 1) r->qs += l, *qshift = l;
+		if ((p->cigar[0]&0xf) == 1) {
-		else r->rs += l, *tshift = l;
+			if (r->rev) r->qe -= l;
+			else r->qs += l;
+			*qshift = l;
+		} else r->rs += l, *tshift = l;
 		--p->n_cigar;
 		memmove(p->cigar, p->cigar + 1, p->n_cigar * 4);
 	}
 }
 
-static void mm_update_extra(mm_reg1_t *r, const uint8_t *qseq, const uint8_t *qual, const uint8_t *tseq, const int8_t *mat, int8_t q, int8_t e)
+static void mm_update_extra(mm_reg1_t *r, const uint8_t *qseq, const uint8_t *tseq, const int8_t *mat, int8_t q, int8_t e)
 {
 	uint32_t k, l, toff = 0, qoff = 0;
 	int32_t s = 0, max = 0, qshift, tshift;
@@ -219,7 +223,7 @@ static inline int mm_get_hplen_back(const mm_idx_t *mi, uint32_t rid, uint32_t x
 
 static inline void mm_adjust_minier(const mm_idx_t *mi, uint8_t *const qseq0[2], mm128_t *a, int32_t *r, int32_t *q)
 {
-	if (mi->is_hpc) {
+	if (mi->flag & MM_I_HPC) {
 		const uint8_t *qseq = qseq0[a->x>>63];
 		int i, c;
 		*q = (int32_t)a->y;
@@ -338,7 +342,49 @@ static void mm_max_stretch(const mm_mapopt_t *opt, const mm_reg1_t *r, const mm1
 	*as = max_i, *cnt = max_len;
 }
 
-static void mm_align1(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, uint8_t *qseq0[2], uint8_t *qual0[2], mm_reg1_t *r, mm_reg1_t *r2, int n_a, mm128_t *a, ksw_extz_t *ez, int splice_flag)
+static int mm_seed_ext_score(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, const int8_t mat[25], int qlen, uint8_t *qseq0[2], const mm128_t *a)
+{
+	uint8_t *qseq, *tseq;
+	int q_span = a->y>>32&0xff, qs, qe, rs, re, rid, score, q_off, t_off, ext_len = opt->anchor_ext_len;
+	void *qp;
+	rid = a->x<<1>>33;
+	re = (uint32_t)a->x + 1, rs = re - q_span;
+	qe = (uint32_t)a->y + 1, qs = qe - q_span;
+	rs = rs - ext_len > 0? rs - ext_len : 0;
+	qs = qs - ext_len > 0? qs - ext_len : 0;
+	re = re + ext_len < mi->seq[rid].len? re + ext_len : mi->seq[rid].len;
+	qe = qe + ext_len < qlen? qe + ext_len : qlen;
+	tseq = (uint8_t*)kmalloc(km, re - rs);
+	mm_idx_getseq(mi, rid, rs, re, tseq);
+	qseq = qseq0[a->x>>63] + qs;
+	qp = ksw_ll_qinit(km, 2, qe - qs, qseq, 5, mat);
+	score = ksw_ll_i16(qp, re - rs, tseq, opt->q, opt->e, &q_off, &t_off);
+	kfree(km, tseq);
+	kfree(km, qp);
+	return score;
+}
+
+static void mm_fix_bad_ends_splice(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, const mm_reg1_t *r, const int8_t mat[25], int qlen, uint8_t *qseq0[2], const mm128_t *a, int *as1, int *cnt1)
+{ // this assumes a very crude k-mer based mode; it is not necessary to use a good model just for filtering bounary exons
+	int score;
+	double log_gap;
+	*as1 = r->as, *cnt1 = r->cnt;
+	if (r->cnt < 3) return;
+	log_gap = log((int32_t)a[r->as + 1].x - (int32_t)a[r->as].x);
+	if ((a[r->as].y>>32&0xff) < log_gap + opt->anchor_ext_shift) {
+		score = mm_seed_ext_score(km, opt, mi, mat, qlen, qseq0, &a[r->as]);
+		if ((double)score / mat[0] < log_gap + opt->anchor_ext_shift) // a more exact format is "score < log_4(gap) + shift"
+			++(*as1), --(*cnt1);
+	}
+	log_gap = log((int32_t)a[r->as + r->cnt - 1].x - (int32_t)a[r->as + r->cnt - 2].x);
+	if ((a[r->as + r->cnt - 1].y>>32&0xff) < log_gap + opt->anchor_ext_shift) {
+		score = mm_seed_ext_score(km, opt, mi, mat, qlen, qseq0, &a[r->as + r->cnt - 1]);
+		if ((double)score / mat[0] < log_gap + opt->anchor_ext_shift)
+			--(*cnt1);
+	}
+}
+
+static void mm_align1(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, uint8_t *qseq0[2], mm_reg1_t *r, mm_reg1_t *r2, int n_a, mm128_t *a, ksw_extz_t *ez, int splice_flag)
 {
 	int is_sr = !!(opt->flag & MM_F_SR), is_splice = !!(opt->flag & MM_F_SPLICE);
 	int32_t rid = a[r->as].x<<1>>33, rev = a[r->as].x>>63, as1, cnt1;
@@ -348,23 +394,25 @@ static void mm_align1(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int
 	int32_t rs1, qs1, re1, qe1;
 	int8_t mat[25];
 
-	if (is_sr) assert(!mi->is_hpc); // HPC won't work with SR because with HPC we can't easily tell if there is a gap
+	if (is_sr) assert(!(mi->flag & MM_I_HPC)); // HPC won't work with SR because with HPC we can't easily tell if there is a gap
 
 	r2->cnt = 0;
 	if (r->cnt == 0) return;
 	ksw_gen_simple_mat(5, mat, opt->a, opt->b);
 	bw = (int)(opt->bw * 1.5 + 1.);
 
-	if (is_sr && !mi->is_hpc) {
+	if (is_sr && !(mi->flag & MM_I_HPC)) {
 		mm_max_stretch(opt, r, a, &as1, &cnt1);
 		rs = (int32_t)a[as1].x + 1 - (int32_t)(a[as1].y>>32&0xff);
 		qs = (int32_t)a[as1].y + 1 - (int32_t)(a[as1].y>>32&0xff);
 		re = (int32_t)a[as1+cnt1-1].x + 1;
 		qe = (int32_t)a[as1+cnt1-1].y + 1;
 	} else {
-		if (!is_splice)
+		if (is_splice) {
+			mm_fix_bad_ends_splice(km, opt, mi, r, mat, qlen, qseq0, a, &as1, &cnt1);
+		} else {
 			mm_fix_bad_ends(r, a, opt->bw, &as1, &cnt1);
-		else as1 = r->as, cnt1 = r->cnt;
+		}
 		mm_filter_bad_seeds(km, as1, cnt1, a, 10, 40, opt->max_gap>>1, 10);
 		mm_adjust_minier(mi, qseq0, &a[as1], &rs, &qs);
 		mm_adjust_minier(mi, qseq0, &a[as1 + cnt1 - 1], &re, &qe);
@@ -467,7 +515,7 @@ static void mm_align1(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int
 
 	for (i = is_sr? cnt1 - 1 : 1; i < cnt1; ++i) { // gap filling
 		if ((a[as1+i].y & (MM_SEED_IGNORE|MM_SEED_TANDEM)) && i != cnt1 - 1) continue;
-		if (is_sr && !mi->is_hpc) {
+		if (is_sr && !(mi->flag & MM_I_HPC)) {
 			re = (int32_t)a[as1 + i].x + 1;
 			qe = (int32_t)a[as1 + i].y + 1;
 		} else mm_adjust_minier(mi, qseq0, &a[as1 + i], &re, &qe);
@@ -530,7 +578,7 @@ static void mm_align1(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int
 	assert(re1 - rs1 <= re0 - rs0);
 	if (r->p) {
 		mm_idx_getseq(mi, rid, rs1, re1, tseq);
-		mm_update_extra(r, &qseq0[r->rev][qs1], qual0[r->rev]? &qual0[r->rev][qs1] : 0, tseq, mat, opt->q, opt->e);
+		mm_update_extra(r, &qseq0[r->rev][qs1], tseq, mat, opt->q, opt->e);
 		if (rev && r->p->trans_strand)
 			r->p->trans_strand ^= 3; // flip to the read strand
 	}
@@ -538,10 +586,10 @@ static void mm_align1(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int
 	kfree(km, tseq);
 }
 
-static int mm_align1_inv(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, uint8_t *qseq0[2], uint8_t *qual0[2], const mm_reg1_t *r1, const mm_reg1_t *r2, mm_reg1_t *r_inv, ksw_extz_t *ez)
+static int mm_align1_inv(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, uint8_t *qseq0[2], const mm_reg1_t *r1, const mm_reg1_t *r2, mm_reg1_t *r_inv, ksw_extz_t *ez)
 {
 	int tl, ql, score, ret = 0, q_off, t_off;
-	uint8_t *tseq, *qseq, *qual;
+	uint8_t *tseq, *qseq;
 	int8_t mat[25];
 	void *qp;
 
@@ -559,7 +607,6 @@ static int mm_align1_inv(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, i
 	tseq = (uint8_t*)kmalloc(km, tl);
 	mm_idx_getseq(mi, r1->rid, r1->re, r2->rs, tseq);
 	qseq = &qseq0[!r1->rev][qlen - r2->qs];
-	qual = qual0[!r1->rev]? &qseq0[!r1->rev][qlen - r2->qs] : 0;
 
 	mm_seq_rev(ql, qseq);
 	mm_seq_rev(tl, tseq);
@@ -579,9 +626,10 @@ static int mm_align1_inv(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, i
 	r_inv->inv = 1;
 	r_inv->rev = !r1->rev;
 	r_inv->rid = r1->rid;
+	r_inv->div = -1.0f;
 	r_inv->qs = r1->qe + q_off, r_inv->qe = r_inv->qs + ez->max_q + 1;
 	r_inv->rs = r1->re + t_off, r_inv->re = r_inv->rs + ez->max_t + 1;
-	mm_update_extra(r_inv, &qseq[q_off], qual? &qual[q_off] : 0, &tseq[t_off], mat, opt->q, opt->e);
+	mm_update_extra(r_inv, &qseq[q_off], &tseq[t_off], mat, opt->q, opt->e);
 	ret = 1;
 end_align1_inv:
 	kfree(km, tseq);
@@ -598,11 +646,11 @@ static inline mm_reg1_t *mm_insert_reg(const mm_reg1_t *r, int i, int *n_regs, m
 	return regs;
 }
 
-mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, const char *qstr, const char *qual, int *n_regs_, mm_reg1_t *regs, mm128_t *a)
+mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, const char *qstr, int *n_regs_, mm_reg1_t *regs, mm128_t *a)
 {
 	extern unsigned char seq_nt4_table[256];
 	int32_t i, n_regs = *n_regs_, n_a;
-	uint8_t *qseq0[2], *qual0[2];
+	uint8_t *qseq0[2];
 	ksw_extz_t ez;
 
 	// encode the query sequence
@@ -612,12 +660,6 @@ mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *m
 		qseq0[0][i] = seq_nt4_table[(uint8_t)qstr[i]];
 		qseq0[1][qlen - 1 - i] = qseq0[0][i] < 4? 3 - qseq0[0][i] : 4;
 	}
-	if (qual) {
-		qual0[0] = (uint8_t*)kmalloc(km, qlen * 2);
-		qual0[1] = qual0[0] + qlen;
-		for (i = 0; i < qlen; ++i)
-			qual0[0][i] = qual0[1][qlen - 1 - i] = qual[i] - 33;
-	} else qual0[0] = qual0[1] = 0;
 
 	// align through seed hits
 	n_a = mm_squeeze_a(km, n_regs, regs, a);
@@ -628,8 +670,8 @@ mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *m
 			mm_reg1_t s[2], s2[2];
 			int which, trans_strand;
 			s[0] = s[1] = regs[i];
-			mm_align1(km, opt, mi, qlen, qseq0, qual0, &s[0], &s2[0], n_a, a, &ez, MM_F_SPLICE_FOR);
+			mm_align1(km, opt, mi, qlen, qseq0, &s[0], &s2[0], n_a, a, &ez, MM_F_SPLICE_FOR);
-			mm_align1(km, opt, mi, qlen, qseq0, qual0, &s[1], &s2[1], n_a, a, &ez, MM_F_SPLICE_REV);
+			mm_align1(km, opt, mi, qlen, qseq0, &s[1], &s2[1], n_a, a, &ez, MM_F_SPLICE_REV);
 			if (s[0].p->dp_score > s[1].p->dp_score) which = 0, trans_strand = 1;
 			else if (s[0].p->dp_score < s[1].p->dp_score) which = 1, trans_strand = 2;
 			else trans_strand = 3, which = (qlen + s[0].p->dp_score) & 1; // randomly choose a strand, effectively
@@ -642,13 +684,13 @@ mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *m
 			}
 			regs[i].p->trans_strand = trans_strand;
 		} else { // one round of alignment
-			mm_align1(km, opt, mi, qlen, qseq0, qual0, &regs[i], &r2, n_a, a, &ez, opt->flag);
+			mm_align1(km, opt, mi, qlen, qseq0, &regs[i], &r2, n_a, a, &ez, opt->flag);
 			if (opt->flag&MM_F_SPLICE)
 				regs[i].p->trans_strand = opt->flag&MM_F_SPLICE_FOR? 1 : 2;
 		}
 		if (r2.cnt > 0) regs = mm_insert_reg(&r2, i, &n_regs, regs);
 		if (!(opt->flag&MM_F_SPLICE) && !(opt->flag&MM_F_SR) && i > 0) { // don't try inversion alignment for -xsplice or -xsr
-			if (mm_align1_inv(km, opt, mi, qlen, qseq0, qual0, &regs[i-1], &regs[i], &r2, &ez)) {
+			if (mm_align1_inv(km, opt, mi, qlen, qseq0, &regs[i-1], &regs[i], &r2, &ez)) {
 				regs = mm_insert_reg(&r2, i, &n_regs, regs);
 				++i; // skip the inserted INV alignment
 			}
@@ -656,7 +698,6 @@ mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *m
 	}
 	*n_regs_ = n_regs;
 	kfree(km, qseq0[0]);
-	if (qual0[0]) kfree(km, qual0[0]);
 	kfree(km, ez.cigar);
 	mm_filter_regs(km, opt, n_regs_, regs);
 	mm_hit_sort_by_dp(km, n_regs_, regs);

chain.c

@@ -49,7 +49,7 @@ mm128_t *mm_chain_dp(int max_dist_x, int max_dist_y, int bw, int max_skip, int m
 			int64_t dr = ri - a[j].x;
 			int32_t dq = qi - (int32_t)a[j].y, dd, sc, log_dd;
 			int32_t sidj = (a[j].y & MM_SEED_SEG_MASK) >> MM_SEED_SEG_SHIFT;
-			if (dr == 0 || dq <= 0) continue;
+			if ((sidi == sidj && dr == 0) || dq <= 0) continue; // don't skip if an anchor is used by multiple segments; see below
 			if ((sidi == sidj && dq > max_dist_y) || dq > max_dist_x) continue;
 			dd = dr > dq? dr - dq : dq - dr;
 			if (sidi == sidj && dd > bw) continue;
@@ -61,7 +61,8 @@ mm128_t *mm_chain_dp(int max_dist_x, int max_dist_y, int bw, int max_skip, int m
 				int c_log, c_lin;
 				c_lin = (int)(dd * .01 * avg_qspan);
 				c_log = log_dd;
-				if (dr > dq || sidi != sidj) sc -= c_lin < c_log? c_lin : c_log;
+				if (sidi != sidj && dr == 0) ++sc; // possibly due to overlapping paired ends; give a minor bonus
+				else if (dr > dq || sidi != sidj) sc -= c_lin < c_log? c_lin : c_log;
 				else sc -= c_lin + (c_log>>1);
 			} else sc -= (int)(dd * .01 * avg_qspan) + (log_dd>>1);
 			sc += f[j];

esterr.c

@@ -0,0 +1,64 @@
+#include <math.h>
+#include <stdio.h>
+#include <stdlib.h>
+#include <assert.h>
+#include "mmpriv.h"
+
+static inline int32_t get_for_qpos(int32_t qlen, const mm128_t *a)
+{
+	int32_t x = (int32_t)a->y;
+	int32_t q_span = a->y>>32 & 0xff;
+	if (a->x>>63)
+		x = qlen - 1 - (x + 1 - q_span); // revert the position to the forward strand of query
+	return x;
+}
+
+static int get_mini_idx(int qlen, const mm128_t *a, int32_t n, const uint64_t *mini_pos)
+{
+	int32_t x, L = 0, R = n - 1;
+	x = get_for_qpos(qlen, a);
+	while (L <= R) { // binary search
+		int32_t m = ((uint64_t)L + R) >> 1;
+		int32_t y = (int32_t)mini_pos[m];
+		if (y < x) L = m + 1;
+		else if (y > x) R = m - 1;
+		else return m;
+	}
+	return -1;
+}
+
+void mm_est_err(const mm_idx_t *mi, int qlen, int n_regs, mm_reg1_t *regs, const mm128_t *a, int32_t n, const uint64_t *mini_pos)
+{
+	int i;
+	uint64_t sum_k = 0;
+	float avg_k;
+
+	if (n == 0) return;
+	for (i = 0; i < n; ++i)
+		sum_k += mini_pos[i] >> 32 & 0xff;
+	avg_k = (float)sum_k / n;
+
+	for (i = 0; i < n_regs; ++i) {
+		mm_reg1_t *r = &regs[i];
+		int32_t st, en, j, k, n_match, n_tot, l_ref;
+		r->div = -1.0f;
+		if (r->cnt == 0) continue;
+		st = en = get_mini_idx(qlen, r->rev? &a[r->as + r->cnt - 1] : &a[r->as], n, mini_pos);
+		if (st < 0) {
+			if (mm_verbose >= 2)
+				fprintf(stderr, "[WARNING] logic inconsistency in mm_est_err(). Please contact the developer.\n");
+			continue;
+		}
+		l_ref = mi->seq[r->rid].len;
+		for (k = 1, j = st + 1, n_match = 1; j < n && k < r->cnt; ++j) {
+			int32_t x;
+			x = get_for_qpos(qlen, r->rev? &a[r->as + r->cnt - 1 - k] : &a[r->as + k]);
+			if (x == (int32_t)mini_pos[j])
+				++k, en = j, ++n_match;
+		}
+		n_tot = en - st + 1;
+		if (r->qs > avg_k && r->rs > avg_k) ++n_tot;
+		if (qlen - r->qs > avg_k && l_ref - r->re > avg_k) ++n_tot;
+		r->div = logf((float)n_tot / n_match) / avg_k;
+	}
+}

format.c

@@ -202,8 +202,9 @@ static void write_cs(void *km, kstring_t *s, const mm_idx_t *mi, const mm_bseq1_
 
 static inline void write_tags(kstring_t *s, const mm_reg1_t *r)
 {
-	int type = r->inv? 'I' : r->id == r->parent? 'P' : 'S';
+	int type;
-	if (r->iden_flt) mm_sprintf_lite(s, "\tom:i:%d", r->mapq);
+	if (r->id == r->parent) type = r->inv? 'I' : 'P';
+	else type = r->inv? 'i' : 'S';
 	if (r->p) {
 		mm_sprintf_lite(s, "\tNM:i:%d\tms:i:%d\tAS:i:%d\tnn:i:%d", r->blen - r->mlen + r->p->n_ambi, r->p->dp_max, r->p->dp_score, r->p->n_ambi);
 		if (r->p->trans_strand == 1 || r->p->trans_strand == 2)
@@ -211,6 +212,12 @@ static inline void write_tags(kstring_t *s, const mm_reg1_t *r)
 	}
 	mm_sprintf_lite(s, "\ttp:A:%c\tcm:i:%d\ts1:i:%d", type, r->cnt, r->score);
 	if (r->parent == r->id) mm_sprintf_lite(s, "\ts2:i:%d", r->subsc);
+	if (r->div >= 0.0f && r->div <= 1.0f) {
+		char buf[8];
+		if (r->div == 0.0f) buf[0] = '0', buf[1] = 0;
+		else sprintf(buf, "%.4f", r->div);
+		mm_sprintf_lite(s, "\tdv:f:%s", buf);
+	}
 	if (r->split) mm_sprintf_lite(s, "\tzd:i:%d", r->split);
 }
 
@@ -258,7 +265,7 @@ static inline const mm_reg1_t *get_sam_pri(int n_regs, const mm_reg1_t *regs)
 	return NULL;
 }
 
-static void write_sam_cigar(kstring_t *s, int sam_flag, int in_tag, int qlen, const mm_reg1_t *r)
+static void write_sam_cigar(kstring_t *s, int sam_flag, int in_tag, int qlen, const mm_reg1_t *r, int opt_flag)
 {
 	if (r->p == 0) {
 		mm_sprintf_lite(s, "*");
@@ -267,14 +274,14 @@ static void write_sam_cigar(kstring_t *s, int sam_flag, int in_tag, int qlen, co
 		clip_len[0] = r->rev? qlen - r->qe : r->qs;
 		clip_len[1] = r->rev? r->qs : qlen - r->qe;
 		if (in_tag) {
-			int clip_char = (sam_flag&0x800)? 5 : 4;
+			int clip_char = (sam_flag&0x800) && !(opt_flag&MM_F_SOFTCLIP)? 5 : 4;
 			mm_sprintf_lite(s, "\tCG:B:I");
 			if (clip_len[0]) mm_sprintf_lite(s, ",%u", clip_len[0]<<4|clip_char);
 			for (k = 0; k < r->p->n_cigar; ++k)
 				mm_sprintf_lite(s, ",%u", r->p->cigar[k]);
 			if (clip_len[1]) mm_sprintf_lite(s, ",%u", clip_len[1]<<4|clip_char);
 		} else {
-			int clip_char = (sam_flag&0x800)? 'H' : 'S';
+			int clip_char = (sam_flag&0x800) && !(opt_flag&MM_F_SOFTCLIP)? 'H' : 'S';
 			if (clip_len[0]) mm_sprintf_lite(s, "%d%c", clip_len[0], clip_char);
 			for (k = 0; k < r->p->n_cigar; ++k)
 				mm_sprintf_lite(s, "%d%c", r->p->cigar[k]>>4, "MIDN"[r->p->cigar[k]&0xf]);
@@ -336,9 +343,8 @@ void mm_write_sam2(kstring_t *s, const mm_idx_t *mi, const mm_bseq1_t *t, int se
 			mm_sprintf_lite(s, "\t%s\t%d\t0\t*", mi->seq[this_rid].name, this_pos+1);
 		} else mm_sprintf_lite(s, "\t*\t0\t0\t*");
 	} else {
-		int mapq = !r->iden_flt? r->mapq : r->mapq < 3? r->mapq : 3;
 		this_rid = r->rid, this_pos = r->rs, this_rev = r->rev;
-		mm_sprintf_lite(s, "\t%s\t%d\t%d\t", mi->seq[r->rid].name, r->rs+1, mapq);
+		mm_sprintf_lite(s, "\t%s\t%d\t%d\t", mi->seq[r->rid].name, r->rs+1, r->mapq);
 		if ((opt_flag & MM_F_LONG_CIGAR) && r->p && r->p->n_cigar > max_bam_cigar_op - 2) {
 			int n_cigar = r->p->n_cigar;
 			if (r->qs != 0) ++n_cigar;
@@ -350,7 +356,7 @@ void mm_write_sam2(kstring_t *s, const mm_idx_t *mi, const mm_bseq1_t *t, int se
 			if (flag & 0x100) mm_sprintf_lite(s, "0S"); // secondary alignment
 			else if (flag & 0x800) mm_sprintf_lite(s, "%dS", r->re - r->rs); // supplementary alignment
 			else mm_sprintf_lite(s, "%dS", t->l_seq);
-		} else write_sam_cigar(s, flag, 0, t->l_seq, r);
+		} else write_sam_cigar(s, flag, 0, t->l_seq, r, opt_flag);
 	}
 
 	// write mate positions
@@ -383,7 +389,7 @@ void mm_write_sam2(kstring_t *s, const mm_idx_t *mi, const mm_bseq1_t *t, int se
 		if (t->qual) sam_write_sq(s, t->qual, t->l_seq, 0, 0);
 		else mm_sprintf_lite(s, "*");
 	} else {
-		if ((flag & 0x900) == 0) {
+		if ((flag & 0x900) == 0 || (opt_flag & MM_F_SOFTCLIP)) {
 			sam_write_sq(s, t->seq, t->l_seq, r->rev, r->rev);
 			mm_sprintf_lite(s, "\t");
 			if (t->qual) sam_write_sq(s, t->qual, t->l_seq, r->rev, 0);
@@ -431,7 +437,7 @@ void mm_write_sam2(kstring_t *s, const mm_idx_t *mi, const mm_bseq1_t *t, int se
 		if (r->p && (opt_flag & MM_F_OUT_CS))
 			write_cs(km, s, mi, t, r, !(opt_flag&MM_F_OUT_CS_LONG));
 		if (cigar_in_tag)
-			write_sam_cigar(s, flag, 1, t->l_seq, r);
+			write_sam_cigar(s, flag, 1, t->l_seq, r, opt_flag);
 	}
 
 	s->s[s->l] = 0; // we always have room for an extra byte (see str_enlarge)

hit.c

@@ -76,10 +76,11 @@ mm_reg1_t *mm_gen_regs(void *km, uint32_t hash, int qlen, int n_u, uint64_t *u,
 		mm_reg1_t *ri = &r[i];
 		ri->id = i;
 		ri->parent = MM_PARENT_UNSET;
-		ri->score = z[i].x >> 32;
+		ri->score = ri->score0 = z[i].x >> 32;
 		ri->hash = (uint32_t)z[i].x;
 		ri->cnt = (int32_t)z[i].y;
 		ri->as = z[i].y >> 32;
+		ri->div = -1.0f;
 		mm_reg_set_coor(ri, qlen, a);
 	}
 	kfree(km, z);
@@ -147,7 +148,7 @@ void mm_set_parent(void *km, float mask_level, int n, mm_reg1_t *r, int sub_diff
 				ri->parent = rp->parent;
 				rp->subsc = rp->subsc > ri->score? rp->subsc : ri->score;
 				if (ri->cnt >= rp->cnt) cnt_sub = 1;
-				if (rp->p && ri->p && (rp->rs != ri->rs || rp->re != ri->re || ol != min)) { // the last condition excludes identical hits after DP
+				if (rp->p && ri->p && (rp->rid != ri->rid || rp->rs != ri->rs || rp->re != ri->re || ol != min)) { // the last condition excludes identical hits after DP
 					rp->p->dp_max2 = rp->p->dp_max2 > ri->p->dp_max? rp->p->dp_max2 : ri->p->dp_max;
 					if (rp->p->dp_max - ri->p->dp_max <= sub_diff) cnt_sub = 1;
 				}
@@ -234,7 +235,7 @@ void mm_select_sub(void *km, float pri_ratio, int min_diff, int best_n, int *n_,
 			if (p == i || r[i].inv) { // primary or inversion
 				r[k++] = r[i];
 			} else if ((r[i].score >= r[p].score * pri_ratio || r[i].score + min_diff >= r[p].score) && n_2nd < best_n) {
-				if (!(r[i].qs == r[p].qs && r[i].qe == r[p].qe && r[i].rs == r[p].rs && r[i].re == r[p].re)) // not identical hits
+				if (!(r[i].qs == r[p].qs && r[i].qe == r[p].qe && r[i].rid == r[p].rid && r[i].rs == r[p].rs && r[i].re == r[p].re)) // not identical hits
 					r[k++] = r[i], ++n_2nd;
 				else if (r[i].p) free(r[i].p);
 			} else if (r[i].p) free(r[i].p);
@@ -406,27 +407,33 @@ void mm_seg_free(void *km, int n_segs, mm_seg_t *segs)
 void mm_set_mapq(int n_regs, mm_reg1_t *regs, int min_chain_sc, int match_sc, int rep_len, int is_sr)
 {
 	static const float q_coef = 40.0f;
+	int64_t sum_sc = 0;
+	float uniq_ratio;
 	int i;
+	for (i = 0; i < n_regs; ++i)
+		if (regs[i].parent == regs[i].id)
+			sum_sc += regs[i].score;
+	uniq_ratio = (float)sum_sc / (sum_sc + rep_len);
 	for (i = 0; i < n_regs; ++i) {
 		mm_reg1_t *r = &regs[i];
 		if (r->inv) {
 			r->mapq = 0;
 		} else if (r->parent == r->id) {
 			int mapq, subsc;
-			float pen_s1 = (r->score > 100? 1.0f : 0.01f * r->score) * ((float)r->score / (r->score + rep_len));
+			float pen_s1 = (r->score > 100? 1.0f : 0.01f * r->score) * uniq_ratio;
 			float pen_cm = r->cnt > 10? 1.0f : 0.1f * r->cnt;
 			pen_cm = pen_s1 < pen_cm? pen_s1 : pen_cm;
 			subsc = r->subsc > min_chain_sc? r->subsc : min_chain_sc;
 			if (r->p && r->p->dp_max2 > 0 && r->p->dp_max > 0) {
 				float identity = (float)r->mlen / r->blen;
-				float x = (float)r->p->dp_max2 * subsc / r->p->dp_max / r->score;
+				float x = (float)r->p->dp_max2 * subsc / r->p->dp_max / r->score0;
 				mapq = (int)(identity * pen_cm * q_coef * (1.0f - x * x) * logf((float)r->p->dp_max / match_sc));
 				if (!is_sr) {
 					int mapq_alt = (int)(6.02f * identity * identity * (r->p->dp_max - r->p->dp_max2) / match_sc + .499f); // BWA-MEM like mapQ, mostly for short reads
 					mapq = mapq < mapq_alt? mapq : mapq_alt; // in case the long-read heuristic fails
 				}
 			} else {
-				float x = (float)subsc / r->score;
+				float x = (float)subsc / r->score0;
 				if (r->p) {
 					float identity = (float)r->mlen / r->blen;
 					mapq = (int)(identity * pen_cm * q_coef * (1.0f - x) * logf((float)r->p->dp_max / match_sc));

index.c

@@ -31,19 +31,19 @@ typedef struct mm_idx_bucket_s {
 void mm_idxopt_init(mm_idxopt_t *opt)
 {
 	memset(opt, 0, sizeof(mm_idxopt_t));
-	opt->k = 15, opt->w = 10, opt->is_hpc = 0;
+	opt->k = 15, opt->w = 10, opt->flag = 0;
 	opt->bucket_bits = 14;
 	opt->mini_batch_size = 50000000;
 	opt->batch_size = 4000000000ULL;
 }
 
-mm_idx_t *mm_idx_init(int w, int k, int b, int is_hpc)
+mm_idx_t *mm_idx_init(int w, int k, int b, int flag)
 {
 	mm_idx_t *mi;
 	if (k*2 < b) b = k * 2;
 	if (w < 1) w = 1;
 	mi = (mm_idx_t*)calloc(1, sizeof(mm_idx_t));
-	mi->w = w, mi->k = k, mi->b = b, mi->is_hpc = is_hpc;
+	mi->w = w, mi->k = k, mi->b = b, mi->flag = flag;
 	mi->B = (mm_idx_bucket_t*)calloc(1<<b, sizeof(mm_idx_bucket_t));
 	if (!(mm_dbg_flag & 1)) mi->km = km_init();
 	return mi;
@@ -89,7 +89,7 @@ void mm_idx_stat(const mm_idx_t *mi)
 {
 	int i, n = 0, n1 = 0;
 	uint64_t sum = 0, len = 0;
-	fprintf(stderr, "[M::%s] kmer size: %d; skip: %d; is_HPC: %d; #seq: %d\n", __func__, mi->k, mi->w, mi->is_hpc, mi->n_seq);
+	fprintf(stderr, "[M::%s] kmer size: %d; skip: %d; is_hpc: %d; #seq: %d\n", __func__, mi->k, mi->w, mi->flag&MM_I_HPC, mi->n_seq);
 	for (i = 0; i < mi->n_seq; ++i)
 		len += mi->seq[i].len;
 	for (i = 0; i < 1<<mi->b; ++i)
@@ -214,7 +214,7 @@ static void mm_idx_post(mm_idx_t *mi, int n_threads)
 #include "bseq.h"
 
 typedef struct {
-	int mini_batch_size, keep_name;
+	int mini_batch_size;
 	uint64_t batch_size, sum_len;
 	mm_bseq_file_t *fp;
 	mm_idx_t *mi;
@@ -246,7 +246,6 @@ static void *worker_pipeline(void *shared, int step, void *in)
 		s->seq = mm_bseq_read(p->fp, p->mini_batch_size, 0, &s->n_seq); // read a mini-batch
 		if (s->seq) {
 			uint32_t old_m, m;
-			uint64_t sum_len, old_max_len, max_len;
 			assert((uint64_t)p->mi->n_seq + s->n_seq <= UINT32_MAX); // to prevent integer overflow
 			// make room for p->mi->seq
 			old_m = p->mi->n_seq, m = p->mi->n_seq + s->n_seq;
@@ -254,29 +253,34 @@ static void *worker_pipeline(void *shared, int step, void *in)
 			if (old_m != m)
 				p->mi->seq = (mm_idx_seq_t*)krealloc(p->mi->km, p->mi->seq, m * sizeof(mm_idx_seq_t));
 			// make room for p->mi->S
-			for (i = 0, sum_len = 0; i < s->n_seq; ++i) sum_len += s->seq[i].l_seq;
+			if (!(p->mi->flag & MM_I_NO_SEQ)) {
-			old_max_len = (p->sum_len + 7) / 8;
+				uint64_t sum_len, old_max_len, max_len;
-			max_len = (p->sum_len + sum_len + 7) / 8;
+				for (i = 0, sum_len = 0; i < s->n_seq; ++i) sum_len += s->seq[i].l_seq;
-			kroundup64(old_max_len); kroundup64(max_len);
+				old_max_len = (p->sum_len + 7) / 8;
-			if (old_max_len != max_len) {
+				max_len = (p->sum_len + sum_len + 7) / 8;
-				p->mi->S = (uint32_t*)realloc(p->mi->S, max_len * 4);
+				kroundup64(old_max_len); kroundup64(max_len);
-				memset(&p->mi->S[old_max_len], 0, 4 * (max_len - old_max_len));
+				if (old_max_len != max_len) {
+					p->mi->S = (uint32_t*)realloc(p->mi->S, max_len * 4);
+					memset(&p->mi->S[old_max_len], 0, 4 * (max_len - old_max_len));
+				}
 			}
 			// populate p->mi->seq
 			for (i = 0; i < s->n_seq; ++i) {
 				mm_idx_seq_t *seq = &p->mi->seq[p->mi->n_seq];
 				uint32_t j;
-				if (p->keep_name) {
+				if (!(p->mi->flag & MM_I_NO_NAME)) {
 					seq->name = (char*)kmalloc(p->mi->km, strlen(s->seq[i].name) + 1);
 					strcpy(seq->name, s->seq[i].name);
 				} else seq->name = 0;
 				seq->len = s->seq[i].l_seq;
 				seq->offset = p->sum_len;
 				// copy the sequence
-				for (j = 0; j < seq->len; ++j) { // TODO: this is not the fastest way, but let's first see if speed matters here
+				if (!(p->mi->flag & MM_I_NO_SEQ)) {
-					uint64_t o = p->sum_len + j;
+					for (j = 0; j < seq->len; ++j) { // TODO: this is not the fastest way, but let's first see if speed matters here
-					int c = seq_nt4_table[(uint8_t)s->seq[i].seq[j]];
+						uint64_t o = p->sum_len + j;
-					mm_seq4_set(p->mi->S, o, c);
+						int c = seq_nt4_table[(uint8_t)s->seq[i].seq[j]];
+						mm_seq4_set(p->mi->S, o, c);
+					}
 				}
 				// update p->sum_len and p->mi->n_seq
 				p->sum_len += seq->len;
@@ -288,7 +292,10 @@ static void *worker_pipeline(void *shared, int step, void *in)
         step_t *s = (step_t*)in;
 		for (i = 0; i < s->n_seq; ++i) {
 			mm_bseq1_t *t = &s->seq[i];
-			mm_sketch(0, t->seq, t->l_seq, p->mi->w, p->mi->k, t->rid, p->mi->is_hpc, &s->a);
+			if (t->l_seq > 0)
+				mm_sketch(0, t->seq, t->l_seq, p->mi->w, p->mi->k, t->rid, p->mi->flag&MM_I_HPC, &s->a);
+			else if (mm_verbose >= 2)
+				fprintf(stderr, "[WARNING] the length database sequence '%s' is 0\n", t->name);
 			free(t->seq); free(t->name);
 		}
 		free(s->seq); s->seq = 0;
@@ -301,16 +308,15 @@ static void *worker_pipeline(void *shared, int step, void *in)
     return 0;
 }
 
-mm_idx_t *mm_idx_gen(mm_bseq_file_t *fp, int w, int k, int b, int is_hpc, int mini_batch_size, int n_threads, uint64_t batch_size, int keep_name)
+mm_idx_t *mm_idx_gen(mm_bseq_file_t *fp, int w, int k, int b, int flag, int mini_batch_size, int n_threads, uint64_t batch_size)
 {
 	pipeline_t pl;
 	if (fp == 0 || mm_bseq_eof(fp)) return 0;
 	memset(&pl, 0, sizeof(pipeline_t));
 	pl.mini_batch_size = mini_batch_size < batch_size? mini_batch_size : batch_size;
-	pl.keep_name = keep_name;
 	pl.batch_size = batch_size;
 	pl.fp = fp;
-	pl.mi = mm_idx_init(w, k, b, is_hpc);
+	pl.mi = mm_idx_init(w, k, b, flag);
 
 	kt_pipeline(n_threads < 3? n_threads : 3, worker_pipeline, &pl, 3);
 	if (mm_verbose >= 3)
@@ -323,13 +329,13 @@ mm_idx_t *mm_idx_gen(mm_bseq_file_t *fp, int w, int k, int b, int is_hpc, int mi
 	return pl.mi;
 }
 
-mm_idx_t *mm_idx_build(const char *fn, int w, int k, int is_hpc, int n_threads) // a simpler interface
+mm_idx_t *mm_idx_build(const char *fn, int w, int k, int flag, int n_threads) // a simpler interface
 {
 	mm_bseq_file_t *fp;
 	mm_idx_t *mi;
 	fp = mm_bseq_open(fn);
 	if (fp == 0) return 0;
-	mi = mm_idx_gen(fp, w, k, 14, is_hpc, 1<<18, n_threads, UINT64_MAX, 1);
+	mi = mm_idx_gen(fp, w, k, 14, flag, 1<<18, n_threads, UINT64_MAX);
 	mm_bseq_close(fp);
 	return mi;
 }
@@ -344,7 +350,7 @@ void mm_idx_dump(FILE *fp, const mm_idx_t *mi)
 	uint32_t x[5];
 	int i;
 
-	x[0] = mi->w, x[1] = mi->k, x[2] = mi->b, x[3] = mi->n_seq, x[4] = mi->is_hpc;
+	x[0] = mi->w, x[1] = mi->k, x[2] = mi->b, x[3] = mi->n_seq, x[4] = mi->flag;
 	fwrite(MM_IDX_MAGIC, 1, 4, fp);
 	fwrite(x, 4, 5, fp);
 	for (i = 0; i < mi->n_seq; ++i) {
@@ -371,7 +377,8 @@ void mm_idx_dump(FILE *fp, const mm_idx_t *mi)
 			fwrite(x, 8, 2, fp);
 		}
 	}
-	fwrite(mi->S, 4, (sum_len + 7) / 8, fp);
+	if (!(mi->flag & MM_I_NO_SEQ))
+		fwrite(mi->S, 4, (sum_len + 7) / 8, fp);
 	fflush(fp);
 }
 
@@ -421,8 +428,10 @@ mm_idx_t *mm_idx_load(FILE *fp)
 			kh_val(h, k) = x[1];
 		}
 	}
-	mi->S = (uint32_t*)malloc((sum_len + 7) / 8 * 4);
+	if (!(mi->flag & MM_I_NO_SEQ)) {
-	fread(mi->S, 4, (sum_len + 7) / 8, fp);
+		mi->S = (uint32_t*)malloc((sum_len + 7) / 8 * 4);
+		fread(mi->S, 4, (sum_len + 7) / 8, fp);
+	}
 	return mi;
 }
 
@@ -476,10 +485,10 @@ mm_idx_t *mm_idx_reader_read(mm_idx_reader_t *r, int n_threads)
 	mm_idx_t *mi;
 	if (r->is_idx) {
 		mi = mm_idx_load(r->fp.idx);
-		if (mi && mm_verbose >= 2 && (mi->k != r->opt.k || mi->w != r->opt.w || mi->is_hpc != r->opt.is_hpc))
+		if (mi && mm_verbose >= 2 && (mi->k != r->opt.k || mi->w != r->opt.w || (mi->flag&MM_I_HPC) != (r->opt.flag&MM_I_HPC)))
 			fprintf(stderr, "[WARNING]\033[1;31m Indexing parameters (-k, -w or -H) overridden by parameters used in the prebuilt index.\033[0m\n");
 	} else
-		mi = mm_idx_gen(r->fp.seq, r->opt.w, r->opt.k, r->opt.bucket_bits, r->opt.is_hpc, r->opt.mini_batch_size, n_threads, r->opt.batch_size, 1);
+		mi = mm_idx_gen(r->fp.seq, r->opt.w, r->opt.k, r->opt.bucket_bits, r->opt.flag, r->opt.mini_batch_size, n_threads, r->opt.batch_size);
 	if (mi) {
 		if (r->fp_out) mm_idx_dump(r->fp_out, mi);
 		++r->n_parts;

kalloc.c

@@ -189,6 +189,10 @@ void km_stat(const void *_km, km_stat_t *s)
 			panic("[km_stat] The end of a free block enters another free block.");
 		if (p->ptr == km->loop_head) break;
 	}
-	for (p = km->core_head; p != NULL; p = p->ptr)
+	for (p = km->core_head; p != NULL; p = p->ptr) {
-		++s->n_cores, s->capacity += p->size * sizeof(header_t);
+		size_t size = p->size * sizeof(header_t);
+		++s->n_cores;
+		s->capacity += size;
+		s->largest = s->largest > size? s->largest : size;
+	}
 }

kalloc.h

@@ -8,7 +8,7 @@ extern "C" {
 #endif
 
 typedef struct {
-	size_t capacity, available, n_blocks, n_cores;
+	size_t capacity, available, n_blocks, n_cores, largest;
 } km_stat_t;
 
 void *kmalloc(void *km, size_t size);

main.c

@@ -6,7 +6,7 @@
 #include "mmpriv.h"
 #include "getopt.h"
 
-#define MM_VERSION "2.4-r555"
+#define MM_VERSION "2.6-r623"
 
 #ifdef __linux__
 #include <sys/resource.h>
@@ -43,6 +43,8 @@ static struct option long_options[] = {
 	{ "end-bonus",      required_argument, 0, 0 },
 	{ "no-pairing",     no_argument,       0, 0 },
 	{ "splice-flank",   optional_argument, 0, 0 },
+	{ "idx-no-seq",     no_argument,       0, 0 },
+	{ "end-seed-pen",   required_argument, 0, 0 },   // 21
 	{ "help",           no_argument,       0, 'h' },
 	{ "max-intron-len", required_argument, 0, 'G' },
 	{ "version",        no_argument,       0, 'V' },
@@ -94,7 +96,7 @@ int main(int argc, char *argv[])
 	while ((c = getopt_long(argc, argv, opt_str, long_options, &long_idx)) >= 0) {
 		if (c == 'w') ipt.w = atoi(optarg);
 		else if (c == 'k') ipt.k = atoi(optarg);
-		else if (c == 'H') ipt.is_hpc = 1;
+		else if (c == 'H') ipt.flag |= MM_I_HPC;
 		else if (c == 'd') fnw = optarg; // the above are indexing related options, except -I
 		else if (c == 'r') opt.bw = (int)mm_parse_num(optarg);
 		else if (c == 't') n_threads = atoi(optarg);
@@ -109,6 +111,7 @@ int main(int argc, char *argv[])
 		else if (c == 'X') opt.flag |= MM_F_AVA | MM_F_NO_SELF;
 		else if (c == 'a') opt.flag |= MM_F_OUT_SAM | MM_F_CIGAR;
 		else if (c == 'Q') opt.flag |= MM_F_NO_QUAL;
+		else if (c == 'Y') opt.flag |= MM_F_SOFTCLIP;
 		else if (c == 'L') opt.flag |= MM_F_LONG_CIGAR;
 		else if (c == 'T') opt.sdust_thres = atoi(optarg);
 		else if (c == 'n') opt.min_cnt = atoi(optarg);
@@ -137,6 +140,8 @@ int main(int argc, char *argv[])
 		else if (c == 0 && long_idx ==13) opt.flag |= MM_F_SR; // --sr
 		else if (c == 0 && long_idx ==17) opt.end_bonus = atoi(optarg); // --end-bonus
 		else if (c == 0 && long_idx ==18) opt.flag |= MM_F_INDEPEND_SEG; // --no-pairing
+		else if (c == 0 && long_idx ==20) ipt.flag |= MM_I_NO_SEQ; // --idx-no-seq
+		else if (c == 0 && long_idx ==21) opt.anchor_ext_shift = atoi(optarg); // --end-seed-pen
 		else if (c == 0 && long_idx == 14) { // --frag
 			if (optarg == 0 || strcmp(optarg, "yes") == 0 || strcmp(optarg, "y") == 0)
 				opt.flag |= MM_F_FRAG_MODE;
@@ -195,6 +200,8 @@ int main(int argc, char *argv[])
 		fprintf(stderr, "[ERROR]\033[1;31m --splice and --frag should not be specified at the same time.\033[0m\n");
 		return 1;
 	}
+	if (!fnw && !(opt.flag&MM_F_CIGAR))
+		ipt.flag |= MM_I_NO_SEQ;
 
 	if (argc == optind || fp_help == stdout) {
 		fprintf(fp_help, "Usage: minimap2 [options] <target.fa>|<target.idx> [query.fa] [...]\n");
@@ -232,6 +239,7 @@ int main(int argc, char *argv[])
 		fprintf(fp_help, "    -R STR       SAM read group line in a format like '@RG\\tID:foo\\tSM:bar' []\n");
 		fprintf(fp_help, "    -c           output CIGAR in PAF\n");
 		fprintf(fp_help, "    --cs[=STR]   output the cs tag; STR is 'short' (if absent) or 'long' [none]\n");
+		fprintf(fp_help, "    -Y           use soft clipping for supplementary alignments\n");
 		fprintf(fp_help, "    -t INT       number of threads [%d]\n", n_threads);
 		fprintf(fp_help, "    -K NUM       minibatch size for mapping [500M]\n");
 //		fprintf(fp_help, "    -v INT       verbose level [%d]\n", mm_verbose);
@@ -257,11 +265,18 @@ int main(int argc, char *argv[])
 	}
 	if (!idx_rdr->is_idx && fnw == 0 && argc - optind < 2) {
 		fprintf(stderr, "[ERROR] missing input: please specify a query file to map or option -d to keep the index\n");
+		mm_idx_reader_close(idx_rdr);
 		return 1;
 	}
 	if (opt.best_n == 0 && (opt.flag&MM_F_CIGAR) && mm_verbose >= 2)
 		fprintf(stderr, "[WARNING]\033[1;31m `-N 0' reduces alignment accuracy. Please use --secondary=no to suppress secondary alignments.\033[0m\n");
 	while ((mi = mm_idx_reader_read(idx_rdr, n_threads)) != 0) {
+		if ((opt.flag & MM_F_CIGAR) && (mi->flag & MM_I_NO_SEQ)) {
+			fprintf(stderr, "[ERROR] the prebuilt index doesn't contain sequences.\n");
+			mm_idx_destroy(mi);
+			mm_idx_reader_close(idx_rdr);
+			return 1;
+		}
 		if ((opt.flag & MM_F_OUT_SAM) && idx_rdr->n_parts == 1) {
 			if (mm_idx_reader_eof(idx_rdr)) {
 				mm_write_sam_hdr(mi, rg, MM_VERSION, argc, argv);

map.c

@@ -36,6 +36,7 @@ void mm_mapopt_init(mm_mapopt_t *opt)
 	opt->end_bonus = -1;
 	opt->min_dp_max = opt->min_chain_score * opt->a;
 	opt->min_ksw_len = 200;
+	opt->anchor_ext_len = 20, opt->anchor_ext_shift = 6;
 	opt->mini_batch_size = 500000000;
 
 	opt->pe_ori = 0; // FF
@@ -64,29 +65,29 @@ int mm_set_opt(const char *preset, mm_idxopt_t *io, mm_mapopt_t *mo)
 		mm_idxopt_init(io);
 		mm_mapopt_init(mo);
 	} else if (strcmp(preset, "ava-ont") == 0) {
-		io->is_hpc = 0, io->k = 15, io->w = 5;
+		io->flag = 0, io->k = 15, io->w = 5;
 		mo->flag |= MM_F_AVA | MM_F_NO_SELF;
 		mo->min_chain_score = 100, mo->pri_ratio = 0.0f, mo->max_gap = 10000, mo->max_chain_skip = 25;
 	} else if (strcmp(preset, "ava-pb") == 0) {
-		io->is_hpc = 1, io->k = 19, io->w = 5;
+		io->flag |= MM_I_HPC, io->k = 19, io->w = 5;
 		mo->flag |= MM_F_AVA | MM_F_NO_SELF;
 		mo->min_chain_score = 100, mo->pri_ratio = 0.0f, mo->max_gap = 10000, mo->max_chain_skip = 25;
 	} else if (strcmp(preset, "map10k") == 0 || strcmp(preset, "map-pb") == 0) {
-		io->is_hpc = 1, io->k = 19;
+		io->flag |= MM_I_HPC, io->k = 19;
 	} else if (strcmp(preset, "map-ont") == 0) {
-		io->is_hpc = 0, io->k = 15;
+		io->flag = 0, io->k = 15;
 	} else if (strcmp(preset, "asm5") == 0) {
-		io->is_hpc = 0, io->k = 19, io->w = 19;
+		io->flag = 0, io->k = 19, io->w = 19;
 		mo->a = 1, mo->b = 19, mo->q = 39, mo->q2 = 81, mo->e = 3, mo->e2 = 1, mo->zdrop = 200;
 		mo->min_dp_max = 200;
 		mo->best_n = 50;
 	} else if (strcmp(preset, "asm10") == 0) {
-		io->is_hpc = 0, io->k = 19, io->w = 19;
+		io->flag = 0, io->k = 19, io->w = 19;
 		mo->a = 1, mo->b = 9, mo->q = 16, mo->q2 = 41, mo->e = 2, mo->e2 = 1, mo->zdrop = 200;
 		mo->min_dp_max = 200;
 		mo->best_n = 50;
 	} else if (strcmp(preset, "short") == 0 || strcmp(preset, "sr") == 0) {
-		io->is_hpc = 0, io->k = 21, io->w = 11;
+		io->flag = 0, io->k = 21, io->w = 11;
 		mo->flag |= MM_F_SR | MM_F_FRAG_MODE | MM_F_NO_PRINT_2ND | MM_F_2_IO_THREADS;
 		mo->pe_ori = 0<<1|1; // FR
 		mo->a = 2, mo->b = 8, mo->q = 12, mo->e = 2, mo->q2 = 24, mo->e2 = 1;
@@ -104,7 +105,7 @@ int mm_set_opt(const char *preset, mm_idxopt_t *io, mm_mapopt_t *mo)
 		mo->max_occ = 5000;
 		mo->mini_batch_size = 50000000;
 	} else if (strcmp(preset, "splice") == 0 || strcmp(preset, "cdna") == 0) {
-		io->is_hpc = 0, io->k = 15, io->w = 5;
+		io->flag = 0, io->k = 15, io->w = 5;
 		mo->flag |= MM_F_SPLICE | MM_F_SPLICE_FOR | MM_F_SPLICE_REV | MM_F_SPLICE_FLANK;
 		mo->max_gap = 2000, mo->max_gap_ref = mo->bw = 200000;
 		mo->a = 1, mo->b = 2, mo->q = 2, mo->e = 1, mo->q2 = 32, mo->e2 = 0;
@@ -122,8 +123,6 @@ typedef struct {
 } mm_match_t;
 
 struct mm_tbuf_s {
-	sdust_buf_t *sdb;
-	mm128_v mini;
 	void *km;
 };
 
@@ -132,24 +131,23 @@ mm_tbuf_t *mm_tbuf_init(void)
 	mm_tbuf_t *b;
 	b = (mm_tbuf_t*)calloc(1, sizeof(mm_tbuf_t));
 	if (!(mm_dbg_flag & 1)) b->km = km_init();
-	b->sdb = sdust_buf_init(b->km);
 	return b;
 }
 
 void mm_tbuf_destroy(mm_tbuf_t *b)
 {
 	if (b == 0) return;
-	kfree(b->km, b->mini.a);
-	sdust_buf_destroy(b->sdb);
 	km_destroy(b->km);
 	free(b);
 }
 
-static int mm_dust_minier(int n, mm128_t *a, int l_seq, const char *seq, int sdust_thres, sdust_buf_t *sdb)
+static int mm_dust_minier(void *km, int n, mm128_t *a, int l_seq, const char *seq, int sdust_thres)
 {
 	int n_dreg, j, k, u = 0;
 	const uint64_t *dreg;
-	if (sdust_thres <= 0 || sdb == 0) return n;
+	sdust_buf_t *sdb;
+	if (sdust_thres <= 0) return n;
+	sdb = sdust_buf_init(km);
 	dreg = sdust_core((const uint8_t*)seq, l_seq, sdust_thres, 64, &n_dreg, sdb);
 	for (j = k = 0; j < n; ++j) { // squeeze out minimizers that significantly overlap with LCRs
 		int32_t qpos = (uint32_t)a[j].y>>1, span = a[j].x&0xff;
@@ -163,45 +161,49 @@ static int mm_dust_minier(int n, mm128_t *a, int l_seq, const char *seq, int sdu
 				l += ee - ss;
 			}
 			if (l <= span>>1) a[k++] = a[j]; // keep the minimizer if less than half of it falls in masked region
-		}
+		} else a[k++] = a[j];
 	}
+	sdust_buf_destroy(sdb);
 	return k; // the new size
 }
 
-static void collect_minimizers(const mm_mapopt_t *opt, const mm_idx_t *mi, int n_segs, const int *qlens, const char **seqs, mm_tbuf_t *b)
+static void collect_minimizers(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int n_segs, const int *qlens, const char **seqs, mm128_v *mv)
 {
 	int i, j, n, sum = 0;
-	b->mini.n = 0;
+	mv->n = 0;
 	for (i = n = 0; i < n_segs; ++i) {
-		mm_sketch(b->km, seqs[i], qlens[i], mi->w, mi->k, i, mi->is_hpc, &b->mini);
+		mm_sketch(km, seqs[i], qlens[i], mi->w, mi->k, i, mi->flag&MM_I_HPC, mv);
-		for (j = n; j < b->mini.n; ++j)
+		for (j = n; j < mv->n; ++j)
-			b->mini.a[j].y += sum << 1;
+			mv->a[j].y += sum << 1;
 		if (opt->sdust_thres > 0) // mask low-complexity minimizers
-			b->mini.n = n + mm_dust_minier(b->mini.n - n, b->mini.a + n, qlens[i], seqs[i], opt->sdust_thres, b->sdb);
+			mv->n = n + mm_dust_minier(km, mv->n - n, mv->a + n, qlens[i], seqs[i], opt->sdust_thres);
-		sum += qlens[i], n = b->mini.n;
+		sum += qlens[i], n = mv->n;
 	}
 }
 
-static mm128_t *collect_seed_hits(const mm_mapopt_t *opt, int max_occ, const mm_idx_t *mi, const char *qname, int qlen, int64_t *n_a, int *rep_len, mm_tbuf_t *b)
+static mm128_t *collect_seed_hits(void *km, const mm_mapopt_t *opt, int max_occ, const mm_idx_t *mi, const char *qname, const mm128_v *mv, int qlen, int64_t *n_a, int *rep_len,
+								  int *n_mini_pos, uint64_t **mini_pos)
 {
 	int rep_st = 0, rep_en = 0, i;
 	mm_match_t *m;
 	mm128_t *a;
 
-	m = (mm_match_t*)kmalloc(b->km, b->mini.n * sizeof(mm_match_t));
+	*n_mini_pos = 0;
-	for (i = 0; i < b->mini.n; ++i) {
+	*mini_pos = (uint64_t*)kmalloc(km, mv->n * sizeof(uint64_t));
+	m = (mm_match_t*)kmalloc(km, mv->n * sizeof(mm_match_t));
+	for (i = 0; i < mv->n; ++i) {
 		int t;
-		mm128_t *p = &b->mini.a[i];
+		mm128_t *p = &mv->a[i];
 		m[i].qpos = (uint32_t)p->y;
 		m[i].cr = mm_idx_get(mi, p->x>>8, &t);
 		m[i].n = t;
 		m[i].seg_id = p->y >> 32;
 	}
-	for (i = 0, *n_a = 0; i < b->mini.n; ++i) // find the length of a[]
+	for (i = 0, *n_a = 0; i < mv->n; ++i) // find the length of a[]
 		if (m[i].n < max_occ) *n_a += m[i].n;
-	a = (mm128_t*)kmalloc(b->km, *n_a * sizeof(mm128_t));
+	a = (mm128_t*)kmalloc(km, *n_a * sizeof(mm128_t));
-	for (i = *rep_len = 0, *n_a = 0; i < b->mini.n; ++i) {
+	for (i = *rep_len = 0, *n_a = 0; i < mv->n; ++i) {
-		mm128_t *p = &b->mini.a[i];
+		mm128_t *p = &mv->a[i];
 		mm_match_t *q = &m[i];
 		const uint64_t *r = q->cr;
 		int k, q_span = p->x & 0xff, is_tandem = 0;
@@ -213,8 +215,9 @@ static mm128_t *collect_seed_hits(const mm_mapopt_t *opt, int max_occ, const mm_
 			} else rep_en = en;
 			continue;
 		}
-		if (i > 0 && p->x>>8 == b->mini.a[i - 1].x>>8) is_tandem = 1;
+		(*mini_pos)[(*n_mini_pos)++] = (uint64_t)q_span<<32 | q->qpos>>1;
-		if (i < b->mini.n - 1 && p->x>>8 == b->mini.a[i + 1].x>>8) is_tandem = 1;
+		if (i > 0 && p->x>>8 == mv->a[i - 1].x>>8) is_tandem = 1;
+		if (i < mv->n - 1 && p->x>>8 == mv->a[i + 1].x>>8) is_tandem = 1;
 		for (k = 0; k < q->n; ++k) {
 			int32_t rpos = (uint32_t)r[k] >> 1;
 			mm128_t *p;
@@ -240,7 +243,7 @@ static mm128_t *collect_seed_hits(const mm_mapopt_t *opt, int max_occ, const mm_
 		}
 	}
 	*rep_len += rep_en - rep_st;
-	kfree(b->km, m);
+	kfree(km, m);
 	return a;
 }
 
@@ -258,7 +261,7 @@ static void chain_post(const mm_mapopt_t *opt, int max_chain_gap_ref, const mm_i
 static mm_reg1_t *align_regs(const mm_mapopt_t *opt, const mm_idx_t *mi, void *km, int qlen, const char *seq, const char *qual, int *n_regs, mm_reg1_t *regs, mm128_t *a)
 {
 	if (!(opt->flag & MM_F_CIGAR)) return regs;
-	regs = mm_align_skeleton(km, opt, mi, qlen, seq, qual, n_regs, regs, a); // this calls mm_filter_regs()
+	regs = mm_align_skeleton(km, opt, mi, qlen, seq, n_regs, regs, a); // this calls mm_filter_regs()
 	if (!(opt->flag & MM_F_AVA)) {
 		mm_set_parent(km, opt->mask_level, *n_regs, regs, opt->a * 2 + opt->b);
 		mm_select_sub(km, opt->pri_ratio, mi->k*2, opt->best_n, n_regs, regs);
@@ -269,13 +272,15 @@ static mm_reg1_t *align_regs(const mm_mapopt_t *opt, const mm_idx_t *mi, void *k
 
 void mm_map_frag(const mm_idx_t *mi, int n_segs, const int *qlens, const char **seqs, const char **quals, int *n_regs, mm_reg1_t **regs, mm_tbuf_t *b, const mm_mapopt_t *opt, const char *qname)
 {
-	int i, j, rep_len, qlen_sum, n_regs0;
+	int i, j, rep_len, qlen_sum, n_regs0, n_mini_pos;
 	int max_chain_gap_qry, max_chain_gap_ref, is_splice = !!(opt->flag & MM_F_SPLICE), is_sr = !!(opt->flag & MM_F_SR);
 	uint32_t hash;
 	int64_t n_a;
-	uint64_t *u;
+	uint64_t *u, *mini_pos;
 	mm128_t *a;
+	mm128_v mv = {0,0,0};
 	mm_reg1_t *regs0;
+	km_stat_t kmst;
 
 	for (i = 0, qlen_sum = 0; i < n_segs; ++i)
 		qlen_sum += qlens[i], n_regs[i] = 0, regs[i] = 0;
@@ -286,8 +291,8 @@ void mm_map_frag(const mm_idx_t *mi, int n_segs, const int *qlens, const char **
 	hash ^= __ac_Wang_hash(qlen_sum) + __ac_Wang_hash(opt->seed);
 	hash  = __ac_Wang_hash(hash);
 
-	collect_minimizers(opt, mi, n_segs, qlens, seqs, b);
+	collect_minimizers(b->km, opt, mi, n_segs, qlens, seqs, &mv);
-	a = collect_seed_hits(opt, opt->mid_occ, mi, qname, qlen_sum, &n_a, &rep_len, b);
+	a = collect_seed_hits(b->km, opt, opt->mid_occ, mi, qname, &mv, qlen_sum, &n_a, &rep_len, &n_mini_pos, &mini_pos);
 	radix_sort_128x(a, a + n_a);
 
 	if (mm_dbg_flag & MM_DBG_PRINT_SEED) {
@@ -327,7 +332,8 @@ void mm_map_frag(const mm_idx_t *mi, int n_segs, const int *qlens, const char **
 		if (rechain) { // redo chaining with a higher max_occ threshold
 			kfree(b->km, a);
 			kfree(b->km, u);
-			a = collect_seed_hits(opt, opt->max_occ, mi, qname, qlen_sum, &n_a, &rep_len, b);
+			kfree(b->km, mini_pos);
+			a = collect_seed_hits(b->km, opt, opt->max_occ, mi, qname, &mv, qlen_sum, &n_a, &rep_len, &n_mini_pos, &mini_pos);
 			radix_sort_128x(a, a + n_a);
 			a = mm_chain_dp(max_chain_gap_ref, max_chain_gap_qry, opt->bw, opt->max_chain_skip, opt->min_cnt, opt->min_chain_score, is_splice, n_segs, n_a, a, &n_regs0, &u, b->km);
 		}
@@ -342,6 +348,7 @@ void mm_map_frag(const mm_idx_t *mi, int n_segs, const int *qlens, const char **
 						i == regs0[j].as? 0 : ((int32_t)a[i].y - (int32_t)a[i-1].y) - ((int32_t)a[i].x - (int32_t)a[i-1].x));
 
 	chain_post(opt, max_chain_gap_ref, mi, b->km, qlen_sum, n_segs, qlens, &n_regs0, regs0, a);
+	if (!is_sr) mm_est_err(mi, qlen_sum, n_regs0, regs0, a, n_mini_pos, mini_pos);
 
 	if (n_segs == 1) { // uni-segment
 		regs0 = align_regs(opt, mi, b->km, qlens[0], seqs[0], quals? quals[0] : 0, &n_regs0, regs0, a);
@@ -361,8 +368,21 @@ void mm_map_frag(const mm_idx_t *mi, int n_segs, const int *qlens, const char **
 			mm_pair(b->km, max_chain_gap_ref, opt->pe_bonus, opt->a * 2 + opt->b, opt->a, qlens, n_regs, regs); // pairing
 	}
 
+	kfree(b->km, mv.a);
 	kfree(b->km, a);
 	kfree(b->km, u);
+	kfree(b->km, mini_pos);
+
+	if (b->km) {
+		km_stat(b->km, &kmst);
+		if (mm_dbg_flag & MM_DBG_PRINT_QNAME)
+			fprintf(stderr, "QM\t%s\t%d\tcap=%ld,nCore=%ld,largest=%ld\n", qname, qlen_sum, kmst.capacity, kmst.n_cores, kmst.largest);
+		assert(kmst.n_blocks == kmst.n_cores); // otherwise, there is a memory leak
+		if (kmst.largest > 1U<<28) {
+			km_destroy(b->km);
+			b->km = km_init();
+		}
+	}
 }
 
 mm_reg1_t *mm_map(const mm_idx_t *mi, int qlen, const char *seq, int *n_regs, mm_tbuf_t *b, const mm_mapopt_t *opt, const char *qname)
@@ -396,14 +416,13 @@ typedef struct {
 static void worker_for(void *_data, long i, int tid) // kt_for() callback
 {
     step_t *s = (step_t*)_data;
-	int *qlens, j, off = s->seg_off[i], pe_ori = s->p->opt->pe_ori, is_sr = !!(s->p->opt->flag & MM_F_SR);
+	int qlens[MM_MAX_SEG], j, off = s->seg_off[i], pe_ori = s->p->opt->pe_ori, is_sr = !!(s->p->opt->flag & MM_F_SR);
-	const char **qseqs, **quals = 0;
+	const char *qseqs[MM_MAX_SEG], *quals[MM_MAX_SEG];
 	mm_tbuf_t *b = s->buf[tid];
+	assert(s->n_seg[i] <= MM_MAX_SEG);
+	memset(quals, 0, sizeof(char*) * MM_MAX_SEG);
 	if (mm_dbg_flag & MM_DBG_PRINT_QNAME)
-		fprintf(stderr, "QR\t%s\t%d\n", s->seq[off].name, tid);
+		fprintf(stderr, "QR\t%s\t%d\t%d\n", s->seq[off].name, tid, s->seq[off].l_seq);
-	qlens = (int*)kmalloc(b->km, s->n_seg[i] * sizeof(int));
-	qseqs = (const char**)kmalloc(b->km, s->n_seg[i] * sizeof(const char**));
-	quals = (const char**)kmalloc(b->km, s->n_seg[i] * sizeof(const char**));
 	for (j = 0; j < s->n_seg[i]; ++j) {
 		if (s->n_seg[i] == 2 && ((j == 0 && (pe_ori>>1&1)) || (j == 1 && (pe_ori&1))))
 			mm_revcomp_bseq(&s->seq[off + j]);
@@ -429,9 +448,6 @@ static void worker_for(void *_data, long i, int tid) // kt_for() callback
 				r->rev = !r->rev;
 			}
 		}
-	kfree(b->km, qlens);
-	kfree(b->km, qseqs);
-	kfree(b->km, quals);
 }
 
 static void *worker_pipeline(void *shared, int step, void *in)

minimap.h

@@ -24,6 +24,11 @@
 #define MM_F_LONG_CIGAR    0x10000
 #define MM_F_INDEPEND_SEG  0x20000
 #define MM_F_SPLICE_FLANK  0x40000
+#define MM_F_SOFTCLIP      0x80000
+
+#define MM_I_HPC          0x1
+#define MM_I_NO_SEQ       0x2
+#define MM_I_NO_NAME      0x4
 
 #define MM_IDX_MAGIC   "MMI\2"
 
@@ -45,7 +50,7 @@ typedef struct {
 } mm_idx_seq_t;
 
 typedef struct {
-	int32_t b, w, k, is_hpc;
+	int32_t b, w, k, flag;
 	uint32_t n_seq;            // number of reference sequences
 	mm_idx_seq_t *seq;         // sequence name, length and offset
 	uint32_t *S;               // 4-bit packed sequence
@@ -63,23 +68,25 @@ typedef struct {
 } mm_extra_t;
 
 typedef struct {
-	int32_t id;                     // ID for internal uses (see also parent below)
+	int32_t id;             // ID for internal uses (see also parent below)
-	uint32_t cnt:30, rev:1, seg_split:1; // number of minimizers; if on the reverse strand
+	int32_t cnt;            // number of minimizers; if on the reverse strand
-	uint32_t rid:31, inv:1;         // reference index; if this is an alignment from inversion rescue
+	int32_t rid;            // reference index; if this is an alignment from inversion rescue
-	int32_t score;                  // DP alignment score
+	int32_t score;          // DP alignment score
-	int32_t qs, qe, rs, re;         // query start and end; reference start and end
+	int32_t qs, qe, rs, re; // query start and end; reference start and end
-	int32_t parent, subsc;          // parent==id if primary; best alternate mapping score
+	int32_t parent, subsc;  // parent==id if primary; best alternate mapping score
-	int32_t as;                     // offset in the a[] array (for internal uses only)
+	int32_t as;             // offset in the a[] array (for internal uses only)
-	int32_t mlen, blen;             // seeded exact match length; seeded alignment block length
+	int32_t mlen, blen;     // seeded exact match length; seeded alignment block length
-	uint32_t mapq:8, split:2, n_sub:22; // mapQ; split pattern; number of suboptimal mappings
+	int32_t n_sub;          // number of suboptimal mappings
-	uint32_t sam_pri:1, proper_frag:1, iden_flt:1, pe_thru:1, dummy:28;
+	int32_t score0;         // initial chaining score (before chain merging/spliting)
+	uint32_t mapq:8, split:2, rev:1, inv:1, sam_pri:1, proper_frag:1, pe_thru:1, seg_split:1, dummy:16;
 	uint32_t hash;
+	float div;
 	mm_extra_t *p;
 } mm_reg1_t;
 
 // indexing and mapping options
 typedef struct {
-	short k, w, is_hpc, bucket_bits;
+	short k, w, flag, bucket_bits;
 	int mini_batch_size;
 	uint64_t batch_size;
 } mm_idxopt_t;
@@ -109,6 +116,7 @@ typedef struct {
 	int end_bonus;
 	int min_dp_max;  // drop an alignment if the score of the max scoring segment is below this threshold
 	int min_ksw_len;
+	int anchor_ext_len, anchor_ext_shift;
 
 	int pe_ori, pe_bonus;
 
@@ -268,7 +276,7 @@ int mm_map_file_frag(const mm_idx_t *idx, int n_segs, const char **fn, const mm_
 
 // deprecated APIs for backward compatibility
 void mm_mapopt_init(mm_mapopt_t *opt);
-mm_idx_t *mm_idx_build(const char *fn, int w, int k, int is_hpc, int n_threads);
+mm_idx_t *mm_idx_build(const char *fn, int w, int k, int flag, int n_threads);
 
 #ifdef __cplusplus
 }

minimap2.1

@@ -1,4 +1,4 @@
-.TH minimap2 1 "6 November 2017" "minimap2-2.4 (r555)" "Bioinformatics tools"
+.TH minimap2 1 "12 December 2017" "minimap2-2.6 (r623)" "Bioinformatics tools"
 .SH NAME
 .PP
 minimap2 - mapping and alignment between collections of DNA sequences
@@ -99,6 +99,14 @@ multiple times to map it against each batch of target sequences.
 may be ending with k/K/m/M/g/G. NB: mapping quality is incorrect given a
 multi-part index.
 .TP
+.B --idx-no-seq
+Don't store target sequences in the index. It saves disk space and memory but
+the index generated with this option will not work with
+.B -a
+or
+.BR -c .
+When base-level alignment is not requested, this option is automatically applied.
+.TP
 .BI -d \ FILE
 Save the minimizer index of
 .I target.fa
@@ -264,6 +272,18 @@ However, the SIRV control does not honor this trend
 on SIRV data, please add
 .B --splice-flank=no
 to the command line.
+.TP
+.BI --end-seed-pen \ INT
+Drop a terminal anchor if
+.IR s <log( g )+ INT ,
+where
+.I s
+is the local alignment score around the anchor and
+.I g
+the length of the terminal gap in the chain. This option is only effective
+with
+.BR --splice .
+It helps to avoid tiny terminal exons. [6]
 .SS Input/output options
 .TP 10
 .B -a
@@ -302,6 +322,9 @@ is given,
 .I short
 is assumed. [none]
 .TP
+.B -Y
+In SAM output, use soft clipping for supplementary alignments.
+.TP
 .BI --seed \ INT
 Integer seed for randomizing equally best hits. Minimap2 hashes
 .I INT
@@ -458,7 +481,7 @@ cb | cb | cb
 r | c | l .
 Tag	Type	Description
 _
-tp	A	Type of aln: P/primary, S/secondary and I/inversion
+tp	A	Type of aln: P/primary, S/secondary and I,i/inversion
 cm	i	Number of minimizers on the chain
 s1	i	Chaining score
 s2	i	Chaining score of the best secondary chain

misc/intron-eval.js

@@ -161,7 +161,8 @@ for (var tid in tr) {
 	if (anno[t[0]] == null) anno[t[0]] = [];
 	var s = t[4];
 	for (var i = 0; i < s.length - 1; ++i) {
-		if (s[i][1] >= s[i+1][0]) throw Error("ERROR: wrong annotation!");
+		if (s[i][1] >= s[i+1][0])
+			warn("WARNING: incorrect annotation for transcript "+tid+" ("+s[i][1]+" >= "+s[i+1][0]+")")
 		anno[t[0]].push([s[i][1], s[i+1][0]]);
 	}
 }

misc/paf2aln.js

@@ -36,20 +36,21 @@ var getopt = function(args, ostr) {
 	return optopt;
 }
 
-var c, maf_out = false, line_len = 80;
+var c, line_len = 80, fmt = "aln";
-while ((c = getopt(arguments, "ml:")) != null) {
+while ((c = getopt(arguments, "f:l:")) != null) {
-	if (c == 'm') maf_out = true;
+	if (c == 'f') {
-	else if (c == 'l') line_len = parseInt(getopt.arg); // TODO: not implemented yet
+		fmt = getopt.arg;
+		if (fmt != "aln" && fmt != "lastz-cigar" && fmt != "maf")
+			throw Error("format must be one of aln, lastz-cigar and maf");
+	} else if (c == 'l') line_len = parseInt(getopt.arg);
 }
 if (line_len == 0) line_len = 0x7fffffff;
 
 if (getopt.ind == arguments.length) {
-	print("Usage: k8 paf2aln.js [options] <with-cs.paf>");
+	print("Usage: k8 paf2aln.js [options] <in.paf>");
 	print("Options:");
-	print("  -m        MAF output (BLAST-like output by default)");
+	print("  -f STR    output format: aln (BLAST-like), maf or lastz-cigar [aln]");
 	print("  -l INT    line length in BLAST-like output [80]");
-	print("");
-	print("Note: this script only works when minimap2 is run with option '-S'");
 	exit(1);
 }
 
@@ -66,7 +67,7 @@ function padding_str(x, len, right)
 function update_aln(s_ref, s_qry, s_mid, type, seq, slen)
 {
 	var l = type == '*'? 1 : seq.length;
-	if (type == '=') {
+	if (type == '=' || type == ':') {
 		s_ref.set(seq);
 		s_qry.set(seq);
 		s_mid.set(Array(l+1).join("|"));
@@ -96,49 +97,73 @@ function print_aln(rs, qs, strand, slen, elen, s_ref, s_qry, s_mid)
 	var st, en;
 	if (strand == '+') st = qs + slen[1] + 1, en = qs + elen[1];
 	else st = qs - slen[1], en = qs - elen[1] + 1;
-	print(["Qry" + strand + ":", padding_str(st,               10, false), s_qry.toString(), padding_str(en          , 10, true)].join(" "));
+	print(["Qry" + strand + ":", padding_str(st, 10, false), s_qry.toString(), padding_str(en, 10, true)].join(" "));
 }
 
-var s_ref = new Bytes(), s_qry = new Bytes(), s_mid = new Bytes();
+var s_ref = new Bytes(), s_qry = new Bytes(), s_mid = new Bytes(); // these are used to show padded alignment
-var re = /([=\-\+\*])([A-Za-z]+)/g;
+var re_cs = /([:=\-\+\*])(\d+|[A-Za-z]+)/g;
+var re_cg = /(\d+)([MIDNSH])/g;
 
 var buf = new Bytes();
-var file = new File(arguments[getopt.ind]);
+var file = arguments[getopt.ind] == "-"? new File() : new File(arguments[getopt.ind]);
-if (maf_out) print("##maf version=1\n");
+var lineno = 0;
+if (fmt == "maf") print("##maf version=1\n");
 while (file.readline(buf) >= 0) {
 	var m, line = buf.toString();
 	var t = line.split("\t", 12);
-	if ((m = /\tcs:Z:(\S+)/.exec(line)) == null) continue;
+	++lineno;
-	var cs = m[1];
 	s_ref.length = s_qry.length = s_mid.length = 0;
 	var slen = [0, 0], elen = [0, 0];
-	if (maf_out) {
+	if (fmt == "lastz-cigar") { // LASTZ-cigar output
-		while ((m = re.exec(cs)) != null)
+		var cg = (m = /\tcg:Z:(\S+)/.exec(line)) != null? m[1] : null;
-			update_aln(s_ref, s_qry, s_mid, m[1], m[2], elen);
+		if (cg == null) {
-		if (maf_out) {
+			warn("WARNING: converting to LASTZ-cigar format requires the 'cg' tag, which is absent on line " + lineno);
-			var score = (m = /\tAS:i:(\d+)/.exec(line)) != null? parseInt(m[1]) : 0;
+			continue;
-			var len = t[0].length > t[5].length? t[0].length : t[5].length;
-			print("a " + score);
-			print(["s", padding_str(t[5], len, true), padding_str(t[7], 10, false), padding_str(parseInt(t[8]) - parseInt(t[7]), 10, false),
-				   "+", padding_str(t[6], 10, false), s_ref.toString()].join(" "));
-			var qs, qe, ql = parseInt(t[1]);
-			if (t[4] == '+') {
-				qs = parseInt(t[2]);
-				qe = parseInt(t[3]);
-			} else {
-				qs = ql - parseInt(t[3]);
-				qe = ql - parseInt(t[2]);
-			}
-			print(["s", padding_str(t[0], len, true), padding_str(qs, 10, false), padding_str(qe - qs, 10, false),
-				   t[4], padding_str(ql, 10, false), s_qry.toString()].join(" "));
-			print("");
 		}
-	} else {
+		var score = (m = /\tAS:i:(\d+)/.exec(line)) != null? m[1] : 0;
-		line = line.replace(/\tc[sg]:Z:\S+/g, "");
+		var out = ['cigar:', t[0], t[2], t[3], t[4], t[5], t[7], t[8], '+', score];
+		while ((m = re_cg.exec(cg)) != null)
+			out.push(m[2], m[1]);
+		print(out.join(" "));
+	} else if (fmt == "maf") { // MAF output
+		var cs = (m = /\tcs:Z:(\S+)/.exec(line)) != null? m[1] : null;
+		if (cs == null) {
+			warn("WARNING: converting to MAF requires the 'cs' tag, which is absent on line " + lineno);
+			continue;
+		}
+		while ((m = re_cs.exec(cs)) != null) {
+			if (m[1] == ':')
+				throw Error("converting to MAF only works with 'minimap2 --cs=long'");
+			update_aln(s_ref, s_qry, s_mid, m[1], m[2], elen);
+		}
+		var score = (m = /\tAS:i:(\d+)/.exec(line)) != null? parseInt(m[1]) : 0;
+		var len = t[0].length > t[5].length? t[0].length : t[5].length;
+		print("a " + score);
+		print(["s", padding_str(t[5], len, true), padding_str(t[7], 10, false), padding_str(parseInt(t[8]) - parseInt(t[7]), 10, false),
+				"+", padding_str(t[6], 10, false), s_ref.toString()].join(" "));
+		var qs, qe, ql = parseInt(t[1]);
+		if (t[4] == '+') {
+			qs = parseInt(t[2]);
+			qe = parseInt(t[3]);
+		} else {
+			qs = ql - parseInt(t[3]);
+			qe = ql - parseInt(t[2]);
+		}
+		print(["s", padding_str(t[0], len, true), padding_str(qs, 10, false), padding_str(qe - qs, 10, false),
+				t[4], padding_str(ql, 10, false), s_qry.toString()].join(" "));
+		print("");
+	} else { // BLAST-like output
+		var cs = (m = /\tcs:Z:(\S+)/.exec(line)) != null? m[1] : null;
+		if (cs == null) {
+			warn("WARNING: converting to BLAST-like alignment requires the 'cs' tag, which is absent on line " + lineno);
+			continue;
+		}
+		line = line.replace(/\tc[sg]:Z:\S+/g, ""); // get rid of cs or cg tags
 		print('>' + line);
 		var rs = parseInt(t[7]), qs = t[4] == '+'? parseInt(t[2]) : parseInt(t[3]);
 		var n_blocks = 0;
-		while ((m = re.exec(cs)) != null) {
+		while ((m = re_cs.exec(cs)) != null) {
+			if (m[1] == ':') m[2] = Array(parseInt(m[2]) + 1).join("=");
 			var start = 0, rest = m[1] == '*'? 1 : m[2].length;
 			while (rest > 0) {
 				var l_proc;

misc/paf2diff.js

@@ -0,0 +1,188 @@
+var getopt = function(args, ostr) {
+	var oli; // option letter list index
+	if (typeof(getopt.place) == 'undefined')
+		getopt.ind = 0, getopt.arg = null, getopt.place = -1;
+	if (getopt.place == -1) { // update scanning pointer
+		if (getopt.ind >= args.length || args[getopt.ind].charAt(getopt.place = 0) != '-') {
+			getopt.place = -1;
+			return null;
+		}
+		if (getopt.place + 1 < args[getopt.ind].length && args[getopt.ind].charAt(++getopt.place) == '-') { // found "--"
+			++getopt.ind;
+			getopt.place = -1;
+			return null;
+		}
+	}
+	var optopt = args[getopt.ind].charAt(getopt.place++); // character checked for validity
+	if (optopt == ':' || (oli = ostr.indexOf(optopt)) < 0) {
+		if (optopt == '-') return null; //  if the user didn't specify '-' as an option, assume it means null.
+		if (getopt.place < 0) ++getopt.ind;
+		return '?';
+	}
+	if (oli+1 >= ostr.length || ostr.charAt(++oli) != ':') { // don't need argument
+		getopt.arg = null;
+		if (getopt.place < 0 || getopt.place >= args[getopt.ind].length) ++getopt.ind, getopt.place = -1;
+	} else { // need an argument
+		if (getopt.place >= 0 && getopt.place < args[getopt.ind].length)
+			getopt.arg = args[getopt.ind].substr(getopt.place);
+		else if (args.length <= ++getopt.ind) { // no arg
+			getopt.place = -1;
+			if (ostr.length > 0 && ostr.charAt(0) == ':') return ':';
+			return '?';
+		} else getopt.arg = args[getopt.ind]; // white space
+		getopt.place = -1;
+		++getopt.ind;
+	}
+	return optopt;
+}
+
+var re_cs = /([:=*+-])(\d+|[A-Za-z]+)/g;
+var c, min_cov_len = 10000, min_var_len = 50000, gap_thres = 50, min_mapq = 5;
+while ((c = getopt(arguments, "l:L:g:q:")) != null) {
+	if (c == 'l') min_cov_len = parseInt(getopt.arg);
+	else if (c == 'L') min_var_len = parseInt(optarg.arg);
+	else if (c == 'g') gap_thres = parseInt(optarg.arg);
+	else if (c == 'q') min_mapq = parseInt(optarg.arg);
+}
+
+if (arguments.length == getopt.ind) {
+	print("Usage: k8 paf2diff.js [options] <with-cs.paf>");
+	print("Options:");
+	print("  -l INT    min alignment length to compute coverage ["+min_cov_len+"]");
+	print("  -L INT    min alignment length to call variants ["+min_var_len+"]");
+	print("  -q INT    min mapping quality ["+min_mapq+"]");
+	print("  -g INT    short/long gap threshold (for statistics only) ["+gap_thres+"]");
+	exit(1);
+}
+
+var file = new File(arguments[getopt.ind]);
+var buf = new Bytes();
+var tot_len = 0, n_sub = [0, 0, 0], n_ins = [0, 0, 0, 0], n_del = [0, 0, 0, 0];
+
+function count_var(o)
+{
+	if (o[3] > 1) return;
+	if (o[5] == '-' && o[6] == '-') return;
+	if (o[5] == '-') { // insertion
+		var l = o[6].length;
+		if (l == 1) ++n_ins[0];
+		else if (l == 2) ++n_ins[1];
+		else if (l < gap_thres) ++n_ins[2];
+		else ++n_ins[3];
+	} else if (o[6] == '-') { // deletion
+		var l = o[5].length;
+		if (l == 1) ++n_del[0];
+		else if (l == 2) ++n_del[1];
+		else if (l < gap_thres) ++n_del[2];
+		else ++n_del[3];
+	} else {
+		++n_sub[0];
+		var s = o[5] + o[6];
+		if (s == 'ag' || s == 'ga' || s == 'ct' || s == 'tc')
+			++n_sub[1];
+		else ++n_sub[2];
+	}
+}
+
+var a = [], out = [];
+var c1_ctg = null, c1_start = 0, c1_end = 0, c1_counted = false, c1_len = 0;
+while (file.readline(buf) >= 0) {
+	var line = buf.toString();
+	if (!/\ts2:i:/.test(line)) continue; // skip secondary alignments
+	var m, t = line.split("\t", 12);
+	for (var i = 6; i <= 11; ++i)
+		t[i] = parseInt(t[i]);
+	if (t[10] < min_cov_len || t[11] < min_mapq) continue;
+	var ctg = t[5], x = t[7], end = t[8];
+	// compute regions covered by 1 contig
+	if (ctg != c1_ctg || x >= c1_end) {
+		if (c1_counted && c1_end > c1_start) {
+			c1_len += c1_end - c1_start;
+			print('R', c1_ctg, c1_start, c1_end);
+		}
+		c1_ctg = ctg, c1_start = x, c1_end = end;
+		c1_counted = (t[10] >= min_var_len);
+	} else if (end > c1_end) { // overlap
+		if (c1_counted && x > c1_start) {
+			c1_len += x - c1_start;
+			print('R', c1_ctg, c1_start, x);
+		}
+		c1_start = c1_end, c1_end = end;
+		c1_counted = (t[10] >= min_var_len);
+	} else { // contained
+		if (c1_counted && x > c1_start) {
+			c1_len += x - c1_start;
+			print('R', c1_ctg, c1_start, x);
+		}
+		c1_start = end;
+	}
+	// output variants ahead of this alignment
+	while (out.length) {
+		if (out[0][0] != ctg || out[0][2] <= x) {
+			count_var(out[0]);
+			print('V', out[0].join("\t"));
+			out.shift();
+		} else break;
+	}
+	// update coverage
+	for (var i = 0; i < out.length; ++i)
+		if (out[i][1] >= x && out[i][2] <= end)
+			++out[i][3];
+	// drop alignments that don't overlap with the current one
+	var k = 0;
+	for (var i = 0; i < a.length; ++i)
+		if (a[0][0] == ctg && a[0][2] > x)
+			a[k++] = a[i];
+	a.length = k;
+	// core loop
+	if (t[10] >= min_var_len) {
+		if ((m = /\tcs:Z:(\S+)/.exec(line)) == null) continue; // no cs tag
+		var cs = m[1];
+		var blen = 0, n_diff = 0;
+		tot_len += t[10];
+		while ((m = re_cs.exec(cs)) != null) {
+			var cov = 1;
+			if (m[1] == '*' || m[1] == '+' || m[1] == '-')
+				for (var i = 0; i < a.length; ++i)
+					if (a[0][2] > x) ++cov;
+			if (m[1] == '=' || m[1] == ':') {
+				var l = m[1] == '='? m[2].length : parseInt(m[2]);
+				x += l, blen += l;
+			} else if (m[1] == '*') {
+				out.push([t[5], x, x+1, cov, t[11], m[2].charAt(0), m[2].charAt(1)]);
+				++x, ++blen, ++n_diff;
+			} else if (m[1] == '+') {
+				out.push([t[5], x, x, cov, t[11], '-', m[2]]);
+				++blen, ++n_diff;
+			} else if (m[1] == '-') {
+				out.push([t[5], x, x + m[2].length, cov, t[11], m[2], '-']);
+				x += m[2].length, ++blen, ++n_diff;
+			}
+		}
+	}
+	a.push([t[5], t[7], t[8]]);
+}
+if (c1_counted && c1_end > c1_start) {
+	c1_len += c1_end - c1_start;
+	print('R', c1_ctg, c1_start, c1_end);
+}
+while (out.length) {
+	count_var(out[0]);
+	print('V', out[0].join("\t"));
+	out.shift();
+}
+
+//warn(tot_len + " alignment columns considered in calling");
+warn(c1_len + " reference bases covered by exactly one contig");
+warn(n_sub[0] + " substitutions; ts/tv = " + (n_sub[1]/n_sub[2]).toFixed(3));
+warn(n_del[0] + " 1bp deletions");
+warn(n_ins[0] + " 1bp insertions");
+warn(n_del[1] + " 2bp deletions");
+warn(n_ins[1] + " 2bp insertions");
+warn(n_del[2] + " [3,"+gap_thres+") deletions");
+warn(n_ins[2] + " [3,"+gap_thres+") insertions");
+warn(n_del[3] + " >="+gap_thres+" deletions");
+warn(n_ins[3] + " >="+gap_thres+" insertions");
+
+buf.destroy();
+file.close();

misc/splice2bed.js

@@ -0,0 +1,127 @@
+var getopt = function(args, ostr) {
+	var oli; // option letter list index
+	if (typeof(getopt.place) == 'undefined')
+		getopt.ind = 0, getopt.arg = null, getopt.place = -1;
+	if (getopt.place == -1) { // update scanning pointer
+		if (getopt.ind >= args.length || args[getopt.ind].charAt(getopt.place = 0) != '-') {
+			getopt.place = -1;
+			return null;
+		}
+		if (getopt.place + 1 < args[getopt.ind].length && args[getopt.ind].charAt(++getopt.place) == '-') { // found "--"
+			++getopt.ind;
+			getopt.place = -1;
+			return null;
+		}
+	}
+	var optopt = args[getopt.ind].charAt(getopt.place++); // character checked for validity
+	if (optopt == ':' || (oli = ostr.indexOf(optopt)) < 0) {
+		if (optopt == '-') return null; //  if the user didn't specify '-' as an option, assume it means null.
+		if (getopt.place < 0) ++getopt.ind;
+		return '?';
+	}
+	if (oli+1 >= ostr.length || ostr.charAt(++oli) != ':') { // don't need argument
+		getopt.arg = null;
+		if (getopt.place < 0 || getopt.place >= args[getopt.ind].length) ++getopt.ind, getopt.place = -1;
+	} else { // need an argument
+		if (getopt.place >= 0 && getopt.place < args[getopt.ind].length)
+			getopt.arg = args[getopt.ind].substr(getopt.place);
+		else if (args.length <= ++getopt.ind) { // no arg
+			getopt.place = -1;
+			if (ostr.length > 0 && ostr.charAt(0) == ':') return ':';
+			return '?';
+		} else getopt.arg = args[getopt.ind]; // white space
+		getopt.place = -1;
+		++getopt.ind;
+	}
+	return optopt;
+}
+
+var colors = ["0,128,255", "255,0,0", "0,192,0"];
+
+function print_lines(a, fmt) {
+	if (a.length == 0) return;
+	if (fmt == "bed") {
+		var n_pri = 0;
+		for (var i = 0; i < a.length; ++i)
+			if (a[i][8] == 0) ++n_pri;
+		if (n_pri > 1) {
+			for (var i = 0; i < a.length; ++i)
+				if (a[i][8] == 0) a[i][8] = 1;
+		} else if (n_pri == 0) {
+			warn("Warning: " + a[0][3] + " doesn't have a primary alignment");
+		}
+		for (var i = 0; i < a.length; ++i) {
+			a[i][8] = colors[a[i][8]];
+			print(a[i].join("\t"));
+		}
+	}
+	a.length = 0;
+}
+
+function main(args) {
+	var re = /(\d+)([MIDNSH])/g;
+	var c, fmt = "bed";
+	while ((c = getopt(args, "f:")) != null) {
+		if (c == 'f') fmt = getopt.arg;
+	}
+	if (getopt.ind == args.length) {
+		warn("Usage: k8 splice2bed.js <in.paf>");
+		exit(1);
+	}
+
+	var file = new File(args[getopt.ind]);
+	var buf = new Bytes();
+	var a = [];
+	while (file.readline(buf) >= 0) {
+		var line = buf.toString();
+		if (line.charAt(0) == '@') continue; // skip SAM header lines
+		var t = line.split("\t");
+		var is_pri = false, cigar = null, a1;
+		if (a.length && a[0][3] != t[0]) {
+			print_lines(a, fmt);
+			a = [];
+		}
+		if (t.length >= 12 && (t[4] == '+' || t[4] == '-')) {
+			for (var i = 12; i < t.length; ++i) {
+				if (t[i].substr(0, 5) == 'cg:Z:') {
+					cigar = t[i].substr(5);
+				} else if (t[i].substr(0, 5) == 's2:i:') {
+					is_pri = true;
+				}
+			}
+			a1 = [t[5], t[7], t[8], t[0], Math.floor(t[9]/t[10]*1000), t[4]];
+		} else if (t.length >= 10) {
+			var flag = parseInt(t[1]);
+			if ((flag&4) || a[2] == '*') continue;
+			cigar = t[5];
+			is_pri = (flag&0x100)? false : true;
+			a1 = [t[2], parseInt(t[3])-1, null, t[0], 1000, (flag&16)? '-' : '+'];
+		} else {
+			throw Error("unrecognized input format");
+		}
+		if (cigar == null) throw Error("missing CIGAR");
+		var m, x0 = 0, x = 0, bs = [], bl = [];
+		while ((m = re.exec(cigar)) != null) {
+			if (m[2] == 'M' || m[2] == 'D') {
+				x += parseInt(m[1]);
+			} else if (m[2] == 'N') {
+				bs.push(x0);
+				bl.push(x - x0);
+				x += parseInt(m[1]);
+				x0 = x;
+			}
+		}
+		bs.push(x0);
+		bl.push(x - x0);
+		// write the BED12 line
+		if (a1[2] == null) a1[2] = a1[1] + x;
+		a1.push(a1[1], a1[2]); // thick start/end is the same as start/end
+		a1.push(is_pri? 0 : 2, bs.length, bl.join(",")+",", bs.join(",")+",");
+		a.push(a1);
+	}
+	print_lines(a, fmt);
+	buf.destroy();
+	file.close();
+}
+
+main(arguments);

mmpriv.h

@@ -64,7 +64,7 @@ const uint64_t *mm_idx_get(const mm_idx_t *mi, uint64_t minier, int *n);
 int mm_idx_getseq(const mm_idx_t *mi, uint32_t rid, uint32_t st, uint32_t en, uint8_t *seq);
 int32_t mm_idx_cal_max_occ(const mm_idx_t *mi, float f);
 mm128_t *mm_chain_dp(int max_dist_x, int max_dist_y, int bw, int max_skip, int min_cnt, int min_sc, int is_cdna, int n_segs, int64_t n, mm128_t *a, int *n_u_, uint64_t **_u, void *km);
-mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, const char *qstr, const char *qual, int *n_regs_, mm_reg1_t *regs, mm128_t *a);
+mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, const char *qstr, int *n_regs_, mm_reg1_t *regs, mm128_t *a);
 
 mm_reg1_t *mm_gen_regs(void *km, uint32_t hash, int qlen, int n_u, uint64_t *u, mm128_t *a);
 void mm_split_reg(mm_reg1_t *r, mm_reg1_t *r2, int n, int qlen, mm128_t *a);
@@ -79,6 +79,8 @@ void mm_join_long(void *km, const mm_mapopt_t *opt, int qlen, int *n_regs, mm_re
 void mm_hit_sort_by_dp(void *km, int *n_regs, mm_reg1_t *r);
 void mm_set_mapq(int n_regs, mm_reg1_t *regs, int min_chain_sc, int match_sc, int rep_len, int is_sr);
 
+void mm_est_err(const mm_idx_t *mi, int qlen, int n_regs, mm_reg1_t *regs, const mm128_t *a, int32_t n, const uint64_t *mini_pos);
+
 mm_seg_t *mm_seg_gen(void *km, uint32_t hash, int n_segs, const int *qlens, int n_regs0, const mm_reg1_t *regs0, int *n_regs, mm_reg1_t **regs, const mm128_t *a);
 void mm_seg_free(void *km, int n_segs, mm_seg_t *segs);
 void mm_pair(void *km, int max_gap_ref, int dp_bonus, int sub_diff, int match_sc, const int *qlens, int *n_regs, mm_reg1_t **regs);

python/cmappy.pxd

@@ -5,7 +5,7 @@ cdef extern from "minimap.h":
 	# Options
 	#
 	ctypedef struct mm_idxopt_t:
-		short k, w, is_hpc, bucket_bits
+		short k, w, flag, bucket_bits
 		int mini_batch_size
 		uint64_t batch_size
 
@@ -30,6 +30,7 @@ cdef extern from "minimap.h":
 		int end_bonus
 		int min_dp_max
 		int min_ksw_len
+		int anchor_ext_len, anchor_ext_shift
 		int pe_ori, pe_bonus
 		float mid_occ_frac
 		int32_t mid_occ
@@ -51,7 +52,7 @@ cdef extern from "minimap.h":
 		pass
 
 	ctypedef struct mm_idx_t:
-		int32_t b, w, k, is_hpc
+		int32_t b, w, k, flag
 		uint32_t n_seq
 		mm_idx_seq_t *seq
 		uint32_t *S

python/mappy.pyx

@@ -13,7 +13,8 @@ cdef class Alignment:
 	cdef _ctg, _cigar # these are python objects
 
 	def __cinit__(self, ctg, cl, cs, ce, strand, qs, qe, mapq, cigar, is_primary, mlen, blen, NM, trans_strand):
-		self._ctg, self._ctg_len, self._r_st, self._r_en = str(ctg), cl, cs, ce
+		self._ctg = ctg if isinstance(ctg, str) else ctg.decode()
+		self._ctg_len, self._r_st, self._r_en = cl, cs, ce
 		self._strand, self._q_st, self._q_en = strand, qs, qe
 		self._NM, self._mlen, self._blen = NM, mlen, blen
 		self._mapq = mapq
@@ -34,7 +35,7 @@ cdef class Alignment:
 	def r_en(self): return self._r_en
 
 	@property
-	def strand(self): return self.strand
+	def strand(self): return self._strand
 
 	@property
 	def trans_strand(self): return self._trans_strand
@@ -150,9 +151,11 @@ def fastx_read(fn):
 	ks = cmappy.mm_fastx_open(str.encode(fn))
 	if ks is NULL: return None
 	while cmappy.kseq_read(ks) >= 0:
-		if ks.qual.l > 0: qual = str(ks.qual.s)
+		if ks.qual.l > 0: qual = ks.qual.s if isinstance(ks.qual.s, str) else ks.qual.s.decode()
 		else: qual = None
-		yield str(ks.name.s), str(ks.seq.s), qual
+		name = ks.name.s if isinstance(ks.name.s, str) else ks.name.s.decode()
+		seq = ks.seq.s if isinstance(ks.seq.s, str) else ks.seq.s.decode()
+		yield name, seq, qual
 	cmappy.mm_fastx_close(ks)
 
 def verbose(v=None):

sdust.c

@@ -56,6 +56,7 @@ sdust_buf_t *sdust_buf_init(void *km)
 	buf = (sdust_buf_t*)kcalloc(km, 1, sizeof(sdust_buf_t));
 	buf->km = km;
 	buf->w = kdq_init(int, buf->km);
+	kdq_resize(int, buf->w, 8);
 	return buf;
 }
 

setup.py

@@ -23,7 +23,7 @@ def readme():
 
 setup(
 	name = 'mappy',
-	version = '2.4',
+	version = '2.6',
 	url = 'https://github.com/lh3/minimap2',
 	description = 'Minimap2 python binding',
 	long_description = readme(),
@@ -35,7 +35,7 @@ setup(
     ext_modules = [Extension('mappy',
 		sources = [module_src, 'align.c', 'bseq.c', 'chain.c', 'format.c', 'hit.c', 'index.c', 'pe.c',
 				   'ksw2_extd2_sse.c', 'ksw2_exts2_sse.c', 'ksw2_extz2_sse.c', 'ksw2_ll_sse.c',
-				   'kalloc.c', 'kthread.c', 'map.c', 'misc.c', 'sdust.c', 'sketch.c'],
+				   'kalloc.c', 'kthread.c', 'map.c', 'misc.c', 'sdust.c', 'sketch.c', 'esterr.c'],
 		depends = ['minimap.h', 'bseq.h', 'kalloc.h', 'kdq.h', 'khash.h', 'kseq.h', 'ksort.h',
 				   'ksw2.h', 'kthread.h', 'kvec.h', 'mmpriv.h', 'sdust.h',
 				   'python/cmappy.h', 'python/cmappy.pxd'],

sketch.c

@@ -115,12 +115,12 @@ void mm_sketch(void *km, const char *str, int len, int w, int k, uint32_t rid, i
 		buf[buf_pos] = info; // need to do this here as appropriate buf_pos and buf[buf_pos] are needed below
 		if (l == w + k - 1) { // special case for the first window - because identical k-mers are not stored yet
 			for (j = buf_pos + 1; j < w; ++j)
-				if (min.x == buf[j].x && buf[j].y != min.y) kv_push(mm128_t, km, *p, buf[j]);
+				if (min.x == buf[j].x && buf[j].y != min.y && buf[j].y != UINT64_MAX) kv_push(mm128_t, km, *p, buf[j]);
 			for (j = 0; j < buf_pos; ++j)
-				if (min.x == buf[j].x && buf[j].y != min.y) kv_push(mm128_t, km, *p, buf[j]);
+				if (min.x == buf[j].x && buf[j].y != min.y && buf[j].y != UINT64_MAX) kv_push(mm128_t, km, *p, buf[j]);
 		}
 		if (info.x <= min.x) { // a new minimum; then write the old min
-			if (l >= w + k) kv_push(mm128_t, km, *p, min);
+			if (l >= w + k && min.y != UINT64_MAX) kv_push(mm128_t, km, *p, min);
 			min = info, min_pos = buf_pos;
 		} else if (buf_pos == min_pos) { // old min has moved outside the window
 			if (l >= w + k - 1) kv_push(mm128_t, km, *p, min);
@@ -130,9 +130,9 @@ void mm_sketch(void *km, const char *str, int len, int w, int k, uint32_t rid, i
 				if (min.x >= buf[j].x) min = buf[j], min_pos = j;
 			if (l >= w + k - 1) { // write identical k-mers
 				for (j = buf_pos + 1; j < w; ++j) // these two loops make sure the output is sorted
-					if (min.x == buf[j].x && min.y != buf[j].y) kv_push(mm128_t, km, *p, buf[j]);
+					if (min.x == buf[j].x && min.y != buf[j].y && buf[j].y != UINT64_MAX) kv_push(mm128_t, km, *p, buf[j]);
 				for (j = 0; j <= buf_pos; ++j)
-					if (min.x == buf[j].x && min.y != buf[j].y) kv_push(mm128_t, km, *p, buf[j]);
+					if (min.x == buf[j].x && min.y != buf[j].y && buf[j].y != UINT64_MAX) kv_push(mm128_t, km, *p, buf[j]);
 			}
 		}
 		if (++buf_pos == w) buf_pos = 0;