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[![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 | ||
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git clone https://github.com/lh3/bwa.git | ||
cd bwa; make | ||
./bwa index ref.fa | ||
./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 | ||
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##Introduction | ||
## Introduction | ||
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BWA is a software package for mapping DNA sequences against a large reference | ||
genome, such as the human genome. It consists of three algorithms: | ||
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different sub-commands: **aln/samse/sampe** for BWA-backtrack, | ||
**bwasw** for BWA-SW and **mem** for the BWA-MEM algorithm. | ||
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##Availability | ||
## Availability | ||
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BWA is released under [GPLv3][1]. The latest source code is [freely | ||
available at github][2]. Released packages can [be downloaded][3] at | ||
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and 3rd-party tools for proper BAM-to-FASTQ conversion, mapping to ALT contigs, | ||
adapter triming, duplicate marking, HLA typing and associated data files. | ||
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##Seeking helps | ||
## Seeking help | ||
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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 | ||
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@@ -46,7 +46,7 @@ have questions about BWA, you may [sign up the mailing list][6] and then send | |
the questions to [[email protected]][7]. You may also ask questions | ||
in forums such as [BioStar][8] and [SEQanswers][9]. | ||
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##Citing BWA | ||
## Citing BWA | ||
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* Li H. and Durbin R. (2009) Fast and accurate short read alignment with | ||
Burrows-Wheeler transform. *Bioinformatics*, **25**, 1754-1760. [PMID: | ||
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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. | ||
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##Frequently asked questions (FAQs) | ||
## Frequently asked questions (FAQs) | ||
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1. [What types of data does BWA work with?](#type) | ||
2. [Why does a read appear multiple times in the output SAM?](#multihit) | ||
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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) | ||
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####<a name="type"></a>1. What types of data does BWA work with? | ||
#### <a name="type"></a>1. What types of data does BWA work with? | ||
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BWA works with a variety types of DNA sequence data, though the optimal | ||
algorithm and setting may vary. The following list gives the recommended | ||
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As is shown above, with non-default settings, BWA-MEM works with Oxford Nanopore | ||
reads with a sequencing error rate over 20%. | ||
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####<a name="multihit"></a>2. Why does a read appear multiple times in the output SAM? | ||
#### <a name="multihit"></a>2. Why does a read appear multiple times in the output SAM? | ||
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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, | ||
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. | ||
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####<a name="4gb"></a>3. Does BWA work on reference sequences longer than 4GB in total? | ||
#### <a name="4gb"></a>3. Does BWA work on reference sequences longer than 4GB in total? | ||
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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. | ||
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####<a name="pe0"></a>4. Why can one read in a pair has high mapping quality but the other has zero? | ||
#### <a name="pe0"></a>4. Why can one read in a pair have a high mapping quality but the other has zero? | ||
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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. | ||
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####<a name="endref"></a>5. How can a BWA-backtrack alignment stands out of the end of a chromosome? | ||
#### <a name="endref"></a>5. How can a BWA-backtrack alignment stand out of the end of a chromosome? | ||
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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 | ||
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. | ||
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####<a name="altctg"></a>6. Does BWA work with ALT contigs in the GRCh38 release? | ||
#### <a name="altctg"></a>6. Does BWA work with ALT contigs in the GRCh38 release? | ||
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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 | ||
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. | ||
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####<a name="postalt"></a>7. Can I just run BWA-MEM against GRCh38+ALT without post-processing? | ||
#### <a name="postalt"></a>7. Can I just run BWA-MEM against GRCh38+ALT without post-processing? | ||
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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 | ||
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