This doesn't seem to be implemented within mpileup. One could either re-write those lines using a perl or awk script, or else disable split-read alignments during the preceding read mapping.

                                                        -  tom blackwell  -

On Wed, 26 Aug 2015, ��˧ wrote:

Hi, Tom,

Sorry to bother you again. I have another question.
Following is one line from the result file of "samtools mpileup". Is there an option that could 
filter the "reference skip" ('<' or '>') in the depth column?

chr10_C-T       3147419 G       7       <<,><>, ggggggg

For example, this is what I want:
chr10_C-T       3147419 G       2       ,, gg


Thanks!


-----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


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