好吧,这是本周(2016.10.21-28)的学习任务之一:安装bowtie2并学习其使用方法&参数设置
所以,啃文档咯,官方文档Version 2.2.9 http://bowtie-bio.sourceforge.net/bowtie2/manual.shtml
以下是我的整理。我不生产文档,我只是文档的搬运工么么哒~
Bowtie2适合将长度50-1000bp的reads比对到长的参考序列上。Bowtie 2 indexes the genome with an FM Index
(based on the Burrows-Wheeler Transform or BWT) 。输出结果为SAM格式。已集成在很多软件中,如
TopHat(a fast splice junction mapper for RNA-seq reads),
Cufflinks(transcriptome assembly and isoform quantitiation from RNA-seq reads),
Crossbow( cloud-enabled tool for analyzing reseuqncing data),
Myrna(a cloud-enabled tool for aligning RNA-seq reads and measuring differential gene expression)。
Bowtie1和2的区别:Bowtie 2‘s command-line arguments and genome index format are both different from Bowtie 1‘s.
1,bowtie1出现的早,所以对于测序长度在50bp以下的序列效果不错,而bowtie2在长度在50bp以上的更好。
2,Bowtie 2支持有空位的比对Number of gaps and gap lengths are not restricted, except by way of the configurable scoring scheme.
3,Bowtie 2支持局部比对(local, some chars will be omited/trimmed),也可以全局比对(end-to-end, all char participate)
4,Bowtie 2对最长序列没有要求,但是Bowtie 1最长不能超过1000bp。
5. Bowtie 2 allows alignments to [overlap ambiguous characters] (e.g. `N`s) in the reference. Bowtie 1 does not.
6,Bowtie 2不能比对colorspace reads.
7, Bowtie 2‘s paired-end alignment is more flexible. Try to find unpaired alignments for each mate。
8, Bowtie 2 reports a spectrum of mapping qualities, Bowtie 1 reports either 0 or high。
MUMmer: align 2 very large sequences(eg: 2 genomes)
NUCmer, BLAT/BLAST, Bowtie2: sensitive alignment to short ref seq(eg: a bacterial genome)
安装bowtie2: 直接下载bowtie2-2.2.9-linux-x86_64.zip,解压,修改环境路径即可
Scores: 更高分=更相似
--ma :match bonus
--mp :mismatch penalty
--np :penality for having N in either the read or the ref
--rdg :affine read gap penalty
--rfg :affine ref gap penalty
全局比对栗子:默认,高质量位点的mismatch罚分为-6,长度为2的gap罚分为-11(gap open-5, extension-3),如果在长度为50的read中只有这两个问题,则总分为-17。所以,最好的分数是0,指read和ref完全相同。
default min score threshold:
可以用--score-min设置
-0.6-0.6*L(read长度)
局部比对栗子:罚分同上,但每个match获得bonus,+2,则如果是上面情况,则得分为2*49-6-11=81
default min score threshold:
20+8*ln(L)(read长度)