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  • bwa and nanopore reads

    Has anyone used bwa (0.7.12) with the mem -x ont2d option on nanopore 2D reads? FAQ's in bwa doc says "BWM-MEM works with Oxford Nanopore reads with a sequencing error rate over 20%.
    I've been able to create a .sam file with about 40,000 2d reads aligned to a single chromosome .fa file, but then have had issues seeing my data.

  • #2
    It works fine for me. What's your problem?

    For extracting mapped reads, you can use samtools (to find mapped reads only) and awk:

    Code:
    samtools view -F 4 input.bam | grep -v '^@' awk '{print "@"$1"\n"$10"\n+\n"$11}' > mapped.fastq

    Comment


    • #3
      sorry I am new at this. thanks for your help.
      here is what I have done:
      I used bwa to index my reference
      ./bwa index /home/nanopore/chr6.fa

      then I used bwa to align my files into a sam file (all 2d fastq files were cat together into one file)

      ./bwa mem -x ont2d /home/nanopore/chr6.fa /home/nanopore/MAP006_10122015.fastq > /home/nanopore/aln.sam

      I then used samtools to convert the sam file to a bam file:

      ./samtools view -bs aln.sam > aln.bam

      and finally sorted the bam file:

      ./samtools sort aln.bam aln_sort


      So the problem I am having is the sam file seems to be readable (at least in a text editor I can see the sequence info along with a header and some alphanumeric codes before each sequence, but the bam files are all symbols)

      If I try to view the bam file in GenomeBrowse I get an error "Data for this zoom level is unavailable until background computation is complete.

      Am I doing something wrong, or missing a step?

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      • #4
        BAM format is a binary representation so it is not surprising that you can no longer "see" the contents of that file. That part should be ok. You workflow is also proper.

        I suggest that you use IGV to view the data instead of Genomebrowse. Follow the user guide here since you are new to IGV.

        Comment


        • #5
          Or Tablet.

          Comment


          • #6
            Thank you!

            My problem was with either my sorting or indexing. Once I used the tools in IGV to sort and index my sam file I can now view my data.

            Tablet looks good too! Thanks for the link.

            Comment

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