Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • thedamian
    Member
    • Feb 2012
    • 50

    Difference between "bedtools bamtofastq" and Picard's SamToFastq

    Hi,
    I need to assembly some unmapped reads from Tophat2's output "unmapped.bam"
    Firstly I have to extract read pairs from unmapped.bam

    I found two tools for it:
    Code:
    bedtools bamtofastq -i unmapped.bam -fq bedtools_un1.fastq -fq2 bedtools_un2.fastq
    and
    Code:
    java -jar SamToFastq.jar I=unmapped.bam F=picard_un1.fastq F2=picard_un2.fastq VALIDATION_STRINGENCY=SILENT
    results:
    bedtools_un1.fastq has 685 MB
    picard_un1.fastq has 915 MB

    My question is, why the difference in the size of files is so big?
    Shouldn't the result files be the same? (as the inmput "unmapped.bam" is the same)
  • TiborNagy
    Senior Member
    • Mar 2010
    • 329

    #2
    Please take a look inside the files. It should be the same size, but maybe one of the programs write out different header or give full header in the quality section.

    Comment

    • thedamian
      Member
      • Feb 2012
      • 50

      #3
      wc -l picard_un1.fastq
      15367376 picard_un1.fastq

      wc -l bedtools_un1.fastq
      11515208 bedtools_un1.fastq

      headers are the same

      Comment

      • dpryan
        Devon Ryan
        • Jul 2011
        • 3478

        #4
        Tophat will often map only one mate from a pair. In those cases, it's up to the program to determine whether such reads should or should not be written to a file. It's quite likely that SamToFastq outputs orphaned reads whereas bamtofastq doesn't.

        Comment

        Latest Articles

        Collapse

        • SEQadmin2
          Nine Things a Sample Prep Scientist Thinks About Before Sequencing
          by SEQadmin2


          I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

          Here are nine questions we think about, in roughly the order they matter, before...
          06-18-2026, 07:11 AM
        • SEQadmin2
          From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
          by SEQadmin2


          Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


          The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
          ...
          06-02-2026, 10:05 AM

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by SEQadmin2, 06-17-2026, 06:09 AM
        0 responses
        36 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-09-2026, 11:58 AM
        0 responses
        100 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-05-2026, 10:09 AM
        0 responses
        120 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-04-2026, 08:59 AM
        0 responses
        113 views
        0 reactions
        Last Post SEQadmin2  
        Working...