Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • NU_454
    Junior Member
    • Feb 2011
    • 5

    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.
  • gringer
    David Eccles (gringer)
    • May 2011
    • 845

    #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

    • NU_454
      Junior Member
      • Feb 2011
      • 5

      #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?

      Comment

      • GenoMax
        Senior Member
        • Feb 2008
        • 7142

        #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

        • gringer
          David Eccles (gringer)
          • May 2011
          • 845

          #5
          Or Tablet.

          Comment

          • NU_454
            Junior Member
            • Feb 2011
            • 5

            #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

            Latest Articles

            Collapse

            • SEQadmin2
              Advanced Sequencing Platforms Tackle Neuroscience’s Toughest Genomics Problems
              by SEQadmin2



              Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
              ...
              07-09-2026, 11:10 AM
            • SEQadmin2
              Cancer Drug Resistance: The Lingering Barrier to Rising Survival
              by SEQadmin2



              Cancer survival rates have significantly increased in the last few decades in the United States, reaching a combined 70% 5-year survival rate by 2021. Behind this number, there are years of research to find new therapies, drug targets, and early detection methods. But there is one core challenge that keeps slowing down these advances, and it’s about drug resistance.

              There is no single reason why many patients don’t respond to treatment as expected. Cancer is...
              07-08-2026, 05:17 AM
            • GATTACAT
              Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
              by GATTACAT
              Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
              07-01-2026, 11:43 AM

            ad_right_rmr

            Collapse

            News

            Collapse

            Topics Statistics Last Post
            Started by SEQadmin2, 07-09-2026, 10:04 AM
            0 responses
            11 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 07-08-2026, 10:08 AM
            0 responses
            9 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 07-07-2026, 11:05 AM
            0 responses
            17 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 07-02-2026, 11:08 AM
            0 responses
            31 views
            0 reactions
            Last Post SEQadmin2  
            Working...