Problem resolved! Thanks very much!
> -----原始邮件----- > 发件人: "Thomas W. Blackwell" <[email protected]> > 发送时间: 2015-08-20 21:25:48 (星期四) > 收件人: "梁芳" <[email protected]> > 抄送: [email protected], "李茹姣" <[email protected]> > 主题: Re: [Samtools-help] Fw: Re: regarding to the result samtools mpileup > > > Might experiment with adding -A. I observe that one read pair has the > proper pair flag set (99,147) and the other pair has it unset (97,145). > The description for -A in the samtools 1.1 man page reads "Do not skip > anomalous read pairs in variant calling." > > - tom blackwell - > > On Thu, 20 Aug 2015, 温帅 wrote: > > > To whom it may concern, > > > I met a problem when using samtools-1.0 mpileup to deal with a pairend reads > mapping result file in sort bam format. The command I used was: > > > samtools mpileup -B -Q 0 -d 999999999 -f ref_all_C-T_G-A.fa > mapping_ms.sort.bam > mapping_ms.sort.bam.pile > > > Following are two paired reads from file "mapping_ms.sort.bam": > > > seq.5547# 97 chr10_C-T 292795 255 100M = > 294319 1624 > GTGATGAGTTGGAGTTGTATTAGTGTTTTTTATGAGAAGGGAGATTTTGGAAATTTAAGAATGATGATTGAGGTGAGGAAGAGGTAGAATTGAGTATTTT > > gggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggg > AS:i:-12 XN:i:0 XM:i:2 XO:i:0 XG:i:0 NM:i:2 MD:Z:64G26T8 > YS:i:0 YT:Z:DP NH:i:1 > seq.5547# 145 chr10_C-T 294319 255 100M = > 292795 -1624 > TATTTATGATGTTTTAGTTTATTGAAAAAGTTTTTGTGTTAATTTAGATAAAGAAGTTAAGTGTTTTTTTATTAAGAATGTTGTATTGGAGTATTTAGAT > > gggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggg > AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:100 YS:i:-12 > YT:Z:DP NH:i:1 > seq.346110# 99 chr10_C-T 294276 255 100M = > 294316 140 > GAATTAGAGTTTGAAATAGAAGTAGTAAGTTTTAGTTAGGAAATATTTATGATGTTTTAGTTTATTGAAAAAGTTTTTGTGTTAATTTAGATAAAGAAGT > > gggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggg > AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:100 YS:i:0 YT:Z:CP > NH:i:1 > seq.346110# 147 chr10_C-T 294316 255 100M = > 294276 -140 > AAATATTTATGATGTTTTAGTTTATTGAAAAAGTTTTTGTGTTAATTTAGATAAAGAAGTTAAGTGTTTTTTTATTAAGAATGTTGTATTGGAGTATTTA > > gggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggg > AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:100 YS:i:0 YT:Z:CP > NH:i:1 > > > > > This is one line from the result file "mapping_ms.sort.bam.pile": > > > > > chr10_C-T 294382 T 1 , g > > > > > From the bam file we could easily know that for position "294382" the depth > should be 2 insead of 1. What options should I use if I don't want to filter > any reads when generated the result file? > > > > > Thanks very much for your time and I am looking forward to your reply. > > > > > Fang Liang > Beijing Institute of Genomics, Chinese Academy of Science > > > > > > > > > > > > > > > > > > > -----鍘熷閭欢----- > 鍙戜欢浜� "Heng Li" <[email protected]> > 鍙戦�佹椂闂� 2015-08-18 23:07:24 (鏄熸湡浜�) > 鏀朵欢浜� "姊佽姵" <[email protected]> > 鎶勯�� "鏉庤尮濮�" <[email protected]> > 涓婚: Re: regarding to the result samtools mpileup > > > The first read is unpaired. mpileup doesn't use it by default. Please ask > [email protected] if you have further questions. > > > Heng > > > On Aug 18, 2015, at 5:14 AM, 姊佽姵 <[email protected]> wrote: > > > > Dear Dr Li, > > > > I met a problem when using samtools-1.0 mpileup to deal with a mapping > > result file in sort bam format. The command I used is: > > > > Following is the command I used to generate the mpileup file: > > samtools mpileup -B -Q 0 -d 999999999 -f ref_all_C-T_G-A.fa > > mapping_ms.sort.bam > mapping_ms.sort.bam.pile > > > > > > This is part of the result file: > > > > chr10_C-T 288153 A 1 . g > > chr10_C-T 288154 T 1 . g > > chr10_C-T 288155 T 1 . g > > chr10_C-T 288156 A 1 . g > > chr10_C-T 288157 G 1 . g > > > > > > Here are two reads from the file mapping_ms.sort.bam from which we could > > see the depth should be 2 for the above position "288155" instead of depth > > 1. > > > > seq.197749# 153 2 chr10_C-T 288137 255 100M = > > 288137 0 > > GTTTGATGATTTTTTTATTAGAGATGTATGAAGTTTATTTTAAATTGGAAAAAAAAAATATATTATGTATTTAAGAATATGTTTTAAGATAAATTGTTTT > > > > gggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggg > > AS:i:0 XN:i:0 XM:i:0 XO:i:0 XG:i:0 NM:i:0 MD:Z:100YT:Z:UP NH:i:1 > > seq.11071# 99 1 chr10_C-T 288146 255 100M = > > 288392 346 > > TTTTTTTATTAGAGATGTATGAAGTTTATTTTAAATTGGAAAAAAAAAATATAGTATGTATTTAAGAATATGTTTTAAGATAAATTGTTTTTAGATATTT > > > > gggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggggg > > AS:i:-6 XN:i:0 XM:i:1 XO:i:0 XG:i:0 NM:i:1 MD:Z:53T46 > > YS:i:-12 YT:Z:CP NH:i:1 > > > > > > What options should I use if I don't want to filter any reads when > > generated the result file? > > > > > > Thanks for your consideration! > > > > 鏉庡崥澹紝鎮ㄥソ锛屾垜鏄兂璇锋暀鎮ㄥ浣曟墠鑳借mpileup鐨勮緭鍑虹粨鏋滈噷鍚湁杈撳叆鏂囦欢鎵�鏈塺eads鐨勮鐩栧害淇℃伅锛屽氨鏄笉鎯宠繃婊ゆ帀浠讳綍涓�鏉ead. > > > > > > Fang Liang > > Beijing Institute of Genomics, Chinese Academy of Science > > > > ------------------------------------------------------------------------------ _______________________________________________ Samtools-help mailing list [email protected] https://lists.sourceforge.net/lists/listinfo/samtools-help
