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26 changes: 13 additions & 13 deletions README.md
Original file line number Diff line number Diff line change
@@ -1,14 +1,14 @@
[![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
./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

##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:
Expand All @@ -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
Expand All @@ -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
Expand All @@ -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:
Expand All @@ -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)
Expand All @@ -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)

####<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?

BWA works with a variety types of DNA sequence data, though the optimal
algorithm and setting may vary. The following list gives the recommended
Expand Down Expand Up @@ -108,42 +108,42 @@ 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%.

####<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?

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.

####<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?

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.

####<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?

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.

####<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?

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.

####<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?

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.

####<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?

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
Expand Down
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