diff --git a/README.md b/README.md index 9ac49bbe..2d186965 100644 --- a/README.md +++ b/README.md @@ -1,6 +1,6 @@ [![Build Status](https://travis-ci.org/lh3/bwa.svg?branch=dev)](https://travis-ci.org/lh3/bwa) [![Build Status](https://drone.io/github.com/lh3/bwa/status.png)](https://drone.io/github.com/lh3/bwa/latest) -##Getting started +## Getting started git clone https://github.com/lh3/bwa.git cd bwa; make @@ -8,7 +8,7 @@ ./bwa mem ref.fa read-se.fq.gz | gzip -3 > aln-se.sam.gz ./bwa mem ref.fa read1.fq read2.fq | gzip -3 > aln-pe.sam.gz -##Introduction +## Introduction BWA is a software package for mapping DNA sequences against a large reference genome, such as the human genome. It consists of three algorithms: @@ -24,7 +24,7 @@ reference genome (the **index** command). Alignment algorithms are invoked with different sub-commands: **aln/samse/sampe** for BWA-backtrack, **bwasw** for BWA-SW and **mem** for the BWA-MEM algorithm. -##Availability +## Availability BWA is released under [GPLv3][1]. The latest source code is [freely available at github][2]. Released packages can [be downloaded][3] at @@ -37,7 +37,7 @@ In addition to BWA, this self-consistent package also comes with bwa-associated and 3rd-party tools for proper BAM-to-FASTQ conversion, mapping to ALT contigs, adapter triming, duplicate marking, HLA typing and associated data files. -##Seeking helps +## Seeking help The detailed usage is described in the man page available together with the source code. You can use `man ./bwa.1` to view the man page in a terminal. The @@ -46,7 +46,7 @@ have questions about BWA, you may [sign up the mailing list][6] and then send the questions to [bio-bwa-help@sourceforge.net][7]. You may also ask questions in forums such as [BioStar][8] and [SEQanswers][9]. -##Citing BWA +## Citing BWA * Li H. and Durbin R. (2009) Fast and accurate short read alignment with Burrows-Wheeler transform. *Bioinformatics*, **25**, 1754-1760. [PMID: @@ -63,7 +63,7 @@ in forums such as [BioStar][8] and [SEQanswers][9]. Please note that the last reference is a preprint hosted at [arXiv.org][13]. I do not have plan to submit it to a peer-reviewed journal in the near future. -##Frequently asked questions (FAQs) +## Frequently asked questions (FAQs) 1. [What types of data does BWA work with?](#type) 2. [Why does a read appear multiple times in the output SAM?](#multihit) @@ -73,7 +73,7 @@ do not have plan to submit it to a peer-reviewed journal in the near future. 6. [Does BWA work with ALT contigs in the GRCh38 release?](#altctg) 7. [Can I just run BWA-MEM against GRCh38+ALT without post-processing?](#postalt) -####1. What types of data does BWA work with? +#### 1. What types of data does BWA work with? BWA works with a variety types of DNA sequence data, though the optimal algorithm and setting may vary. The following list gives the recommended @@ -108,7 +108,7 @@ errors given longer query sequences as the chance of missing all seeds is small. As is shown above, with non-default settings, BWA-MEM works with Oxford Nanopore reads with a sequencing error rate over 20%. -####2. Why does a read appear multiple times in the output SAM? +#### 2. Why does a read appear multiple times in the output SAM? BWA-SW and BWA-MEM perform local alignments. If there is a translocation, a gene fusion or a long deletion, a read bridging the break point may have two hits, @@ -116,18 +116,18 @@ occupying two lines in the SAM output. With the default setting of BWA-MEM, one and only one line is primary and is soft clipped; other lines are tagged with 0x800 SAM flag (supplementary alignment) and are hard clipped. -####3. Does BWA work on reference sequences longer than 4GB in total? +#### 3. Does BWA work on reference sequences longer than 4GB in total? Yes. Since 0.6.x, all BWA algorithms work with a genome with total length over 4GB. However, individual chromosome should not be longer than 2GB. -####4. Why can one read in a pair has high mapping quality but the other has zero? +#### 4. Why can one read in a pair have a high mapping quality but the other has zero? This is correct. Mapping quality is assigned for individual read, not for a read pair. It is possible that one read can be mapped unambiguously, but its mate falls in a tandem repeat and thus its accurate position cannot be determined. -####5. How can a BWA-backtrack alignment stands out of the end of a chromosome? +#### 5. How can a BWA-backtrack alignment stand out of the end of a chromosome? Internally BWA concatenates all reference sequences into one long sequence. A read may be mapped to the junction of two adjacent reference sequences. In this @@ -135,7 +135,7 @@ case, BWA-backtrack will flag the read as unmapped (0x4), but you will see position, CIGAR and all the tags. A similar issue may occur to BWA-SW alignment as well. BWA-MEM does not have this problem. -####6. Does BWA work with ALT contigs in the GRCh38 release? +#### 6. Does BWA work with ALT contigs in the GRCh38 release? Yes, since 0.7.11, BWA-MEM officially supports mapping to GRCh38+ALT. BWA-backtrack and BWA-SW don't properly support ALT mapping as of now. Please @@ -143,7 +143,7 @@ see [README-alt.md][18] for details. Briefly, it is recommended to use [bwakit][17], the binary release of BWA, for generating the reference genome and for mapping. -####7. Can I just run BWA-MEM against GRCh38+ALT without post-processing? +#### 7. Can I just run BWA-MEM against GRCh38+ALT without post-processing? If you are not interested in hits to ALT contigs, it is okay to run BWA-MEM without post-processing. The alignments produced this way are very close to