Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • SAM to BAM problem

    Parse warning at line 27: mapped sequence without CIGAR
    Parse error at line 27: sequence and quality are inconsistent
    Aborted



    I used BWASW for alignment. Any help ??

    Comment


    • Originally posted by abi View Post
      Parse warning at line 27: mapped sequence without CIGAR
      Parse error at line 27: sequence and quality are inconsistent
      Aborted



      I used BWASW for alignment. Any help ??
      Was the file truncated at that line? Otherwise it'd be helpful to see that line of the SAM file.

      Comment


      • No the file was not truncated at line 27. Here is the line:

        0 chr4 96674301 60 227M * 0 0 TCTTTGAAACCAACGAGAACAAAGACACAACATACCAGAATCTCTGGGACACATTTAAAGCAGTGTGTAGAGGGAAATTTACAGCAGTAAATGCCCACAAGAGAATGCAGGAAAGATCTAAAATTGACACCCTAACATCAAAATTAAAAGAACTAGAGAAGCAAGAGCAATCACATTCAAAAGCTAGCAGAAGGCAAGAAATAACTAAGATCAGAGCAGAACTGAAG * NM:i:0 AS:i:227 XS:i:0


        I am using bwa-0.7.4 for my alignment .

        I must say that I used FASTA file for alignment and not FASTQ file. Here is a sample line from FASTA file:

        > MySpecies 207
        GGAGCCTTAAGGTTTGGTTATCGCCTTGTGTTTGTTTCTGGGGGTATCTGTGGGGTATGTGTTTCTGGCCATGTGTCTGTGTCTGTGTCTCTAGGCTGTCTTCTAGTCTCAGCTTGAGATCCACAGGCTTCAAGAGCTCAAGGGGGGAAAAGCCCAATTGTATATAAATTGTGAATGGGACTGATGCGTATGAGACAGGGAGGGTCT

        Comment


        • I have seen similar issues with BWA 0.7 greater, same command runs absolutely fine with 0.7 version of BWA.

          Comment


          • @abi , lh3, sbaheti, dpryan, Hena and rest

            I have the answer to you. There is a bug in newer versions of BWA. It does not produces right SAM files for those sequences which have their Fasta/Fastq files starting with > and a space. As an example:

            This will not produce right SAM files
            > My sequence

            This will produce right SAM files
            >My sequence


            Notice the difference in space after '>'

            Hope this helps. You can read more about me at www.tinyurl.com/abinarain

            Cheers
            Narain
            Last edited by narain; 06-05-2013, 09:40 AM.

            Comment


            • [samopen] SAM header is present: 84 sequences.
              [sam_read1] reference '121337031' is recognized as '*'.
              Parse warning at line 7932555: mapped sequence without CIGAR
              Parse error at line 7932555: sequence and quality are inconsistent
              Aborted (core dumped)


              I am getting this error... tried the suggestion on changing the MDI in the blast2sam.pl but it is still not working!

              Any suggestions will be appreciated.

              Comment


              • [samopen] SAM header is present: 84 sequences.
                [sam_read1] reference '121337031' is recognized as '*'.
                Parse warning at line 7932555: mapped sequence without CIGAR
                Parse error at line 7932555: sequence and quality are inconsistent
                Aborted (core dumped)

                The suggestion to change MDI in blast2sam pl did not work! Any suggestions will be appreciated.

                Comment


                • Blast2Sam won't work, because it is an outdated script (I have tried it several times without success). Try to align your sequences with bwa/bowtie.

                  Comment

                  Latest Articles

                  Collapse

                  • seqadmin
                    Current Approaches to Protein Sequencing
                    by seqadmin


                    Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
                    04-04-2024, 04:25 PM
                  • seqadmin
                    Strategies for Sequencing Challenging Samples
                    by seqadmin


                    Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
                    03-22-2024, 06:39 AM

                  ad_right_rmr

                  Collapse

                  News

                  Collapse

                  Topics Statistics Last Post
                  Started by seqadmin, 04-11-2024, 12:08 PM
                  0 responses
                  18 views
                  0 likes
                  Last Post seqadmin  
                  Started by seqadmin, 04-10-2024, 10:19 PM
                  0 responses
                  22 views
                  0 likes
                  Last Post seqadmin  
                  Started by seqadmin, 04-10-2024, 09:21 AM
                  0 responses
                  17 views
                  0 likes
                  Last Post seqadmin  
                  Started by seqadmin, 04-04-2024, 09:00 AM
                  0 responses
                  49 views
                  0 likes
                  Last Post seqadmin  
                  Working...
                  X