Bowtie的中文意思是:領(lǐng)結(jié),蝴蝶結(jié)
Bowtie2是比對軟件Bowtie的第二版本,主要改進了支持gap比對。
Bowtie2用戶手冊:
http://bowtie-bio.sourceforge.net/bowtie2/manual.shtml
在看比對結(jié)果前需要了解三個概念:
1. Aligned concordantly 合理比對
主要和比對參數(shù):--fr/--rf/--ff?有關(guān),默認是:--fr
在參數(shù)--fr下:前兩種比對就是合理比對,也即:concordantly,后兩種比對會成為不合理比對,也即:aligned discordantly,如果兩個reads距離太遠也會成為不合理比對。
2. Multiple alignments 多重比對
定義:一對reads/一個pair比對上genome的多個位置,也即:aligned >1 times?或是aligned concordantly >1 times,這對reads/這個pair就會成為多重比對。相對應的就是?unique比對,也即:aligned exactly 1 time?或是:aligned concordantly exactly 1 time。
這里 bowtie2的默認參數(shù)是:
Default mode:?search for multiple alignments, report the best one,也即是:多重比對,只取最好的那個alignment。
3. Bowtie2在雙端比對下:
如果一對reads/一個pair沒有比對上任何地方,也即?aligned concordantly 0 times,那么就將這個pair拆分成兩個single-end reads來進行比對。
官網(wǎng)給出的比對Summary示例如下:
Alignment summary
# 單端比對:
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 times93.77% overall alignment rate
# 翻譯下即:
檢測到20000條reads;
????其中,
20000條reads都是unpaired也即單端數(shù)據(jù);
????其中,
1247條reads一次沒比對上;
18937條reads只比對上一次;
14條reads比對多于一次;
???總共的比對率是:(18739+14)/20000=93.77%
# 雙端比對:
For datasets consisting of pairs, the summary might look like this:
10000reads; of these:10000(100.00%) were paired; of these:650(6.50%) aligned concordantly 0 times8823(88.23%) aligned concordantly exactly 1 time527(5.27%) aligned concordantly >1 times----650pairs aligned concordantly 0 times; of these:34(5.23%) aligned discordantly 1 time----616pairs aligned 0 times concordantly or discordantly; of these:1232mates make up the pairs; of these:660(53.57%) aligned 0 times571(46.35%) aligned exactly 1 time1(0.08%) aligned >1 times96.70%overall alignment rate
# 翻譯下即:
檢測到10000個pairs,也即20000條reads;
????其中,
??????? 10000個pairs都是paired也即雙端數(shù)據(jù);
其中,
??????? 650個pairs沒有合理比對上;
8823個pairs只合理比對上一次;
??? ??? 527個pairs合理比對多于一次;
????在650個pairs沒有合理比對上的pairs中:
??????? 34個pairs可以不合理比對上一次;
????剩下的616個pairs,也即1232條reads中:
660條reads沒有比對上;
571條reads可以比對上一次;
??????? 1條read比對多于1次;
總共的比對率是:(8823*2+527*2+34*2+571+1)/20000=96.7%
歡迎探討交流!