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Bowtie的中文意思是:领结,蝴蝶结
Bowtie2是比对软件Bowtie的第二版本,主要改进了支持gap比对。
Bowtie2用户手册:
http://bowtie-bio.sourceforge.net/bowtie2/manual.shtml
在看比对结果前需要了解三个概念:
1. Aligned concordantly 合理比对
主要和比对参数:--fr/--rf/--ff 有关,默认是:--fr
在参数--fr下:前两种比对就是合理比对,也即:concordantly,后两种比对会成为不合理比对,也即:aligned discordantly,如果两个reads距离太远也会成为不合理比对。
2. Multiple alignments 多重比对
定义:一对reads/一个pair比对上genome的多个位置,也即:aligned >1 times 或是 aligned concordantly >1 times,这对reads/这个pair就会成为多重比对。相对应的就是 unique比对,也即:aligned exactly 1 time 或是:aligned concordantly exactly 1 time。
这里 bowtie2的默认参数是:
3. Bowtie2在双端比对下:
如果一对reads/一个pair没有比对上任何地方,也即 aligned concordantly 0 times,那么就将这个pair拆分成两个single-end reads来进行比对。
官网给出的比对Summary示例如下:
Alignment summary
# 单端比对:
For datasets consisting of unpaired reads, the summary might look like this:
20000 reads; of these:
20000 (100.00%) were unpaired; of these:
1247 (6.24%) aligned 0 times
18739 (93.69%) aligned exactly 1 time
14 (0.07%) aligned >1 times
93.77% overall alignment rate
# 翻译下即:
检测到20000条reads;
其中,
20000条reads都是unpaired也即单端数据;
其中,
1247条reads一次没比对上;
18937条reads只比对上一次;
14条reads比对多于一次;
总共的比对率是:(18739+14)/20000=93.77%
# 双端比对:
For datasets consisting of pairs, the summary might look like this:
10000 reads; of these:
10000 (100.00%) were paired; of these:
650 (6.50%) aligned concordantly 0 times
8823 (88.23%) aligned concordantly exactly 1 time
527 (5.27%) aligned concordantly >1 times
----
650 pairs aligned concordantly 0 times; of these:
34 (5.23%) aligned discordantly 1 time
----
616 pairs aligned 0 times concordantly or discordantly; of these:
1232 mates make up the pairs; of these:
660 (53.57%) aligned 0 times
571 (46.35%) aligned exactly 1 time
1 (0.08%) aligned >1 times
96.70% overall alignment rate
# 翻译下即:
检测到10000个pairs,也即20000条reads;
其中,
10000个pairs都是paired也即双端数据;
其中,
650个pairs没有合理比对上;
8823个pairs只合理比对上一次;
527个pairs合理比对多于一次;
在650个pairs没有合理比对上的pairs中:
34个pairs可以不合理比对上一次;
剩下的616个pairs,也即1232条reads中:
660条reads没有比对上;
571条reads可以比对上一次;
1条read比对多于1次;
总共的比对率是:(8823*2+527*2+34*2+571+1)/20000=96.7%
欢迎探讨交流!
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