Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • newbie BWA alignment question

    another newbie question - I am trying to use BWA to align paired end reads to human ref genome.

    here is the sequence of what i did -
    1) alignment
    bwa align ref fq1 > 1.sai

    2) make sam
    bwi sampe ref 1.sai 2.sai fq1 fq2 > .sam

    3) convert to bam
    samtools view -S -b .sam -o .bam

    4) sort bam
    samtools sort .bam .sorted

    5) index bam
    samtools index .sorted

    6) check stats
    samtools flagstat .bam > .stat

    everything finished running without any error, but when i check the .stat file, this is what i see

    0 + 0 duplicates
    0 + 0 mapped (0.00%:nan%)
    61091524 + 0 paired in sequencing
    30545762 + 0 read1
    30545762 + 0 read2
    0 + 0 properly paired (0.00%:nan%)
    0 + 0 with itself and mate mapped
    0 + 0 singletons (0.00%:nan%)
    0 + 0 with mate mapped to a different chr
    0 + 0 with mate mapped to a different chr (mapQ>=5)

    in every sample, there is very few to no aligned reads. can anyone comment on what could possibly be the issue here and if there is a way to check the output after each stage in this process. thanks.

  • #2
    .sams are human readable. So start by looking at those.

    You did index the genome too, right?

    Comment


    • #3
      Did you align the second set of fastq sequences? All I see is
      bwa align ref fq1 > 1.sai
      but not
      bwa align ref fq2 > 2.sai

      Comment

      Latest Articles

      Collapse

      • seqadmin
        Essential Discoveries and Tools in Epitranscriptomics
        by seqadmin


        The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist on Modified Bases...
        Yesterday, 07:01 AM
      • 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

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, 04-11-2024, 12:08 PM
      0 responses
      51 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      50 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      44 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-04-2024, 09:00 AM
      0 responses
      55 views
      0 likes
      Last Post seqadmin  
      Working...
      X