Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • sorted bam larger than unsorted bam

    Recently i've sorted 2 different alignments and in both cases the sorted bam is ~3x larger in disk size. I did the sort twice with the same result.
    Does anyone have a suspicion of what is going on here?

  • #2
    1, output bam uncompressed? (show the command line pls.)
    2, you are sorting on name or coord? could it be that bgzip block can't compress that hard after the sort? (Very unlikely though.)

    Comment


    • #3
      Here's what i'm running:

      Code:
      time -v samtools sort $mergedBam $sortedBam
      Another run yielded the same result.

      Comment


      • #4
        Do you have a combination of shallow coverage and excessively long read names? That can cause the sorting to be of little benefit to sequence and positional compression while also being detrimental to read name compression.

        Comment


        • #5
          headers aren't anything out of the ordinary. Here are a few entries from the sorted bam:

          Code:
          HWI-ST1063_0137:6:1308:10615:65342#0	161	chr1	765	59	95M6S	=	1019	354	TGACGGACTACATGAGATAGAAGAGAGAATTTTGGGAGCAGAAGATATCATAGAAAACATTGACACAACCTTCAAAGAGAACGTAAATAGGAAAAAGCTCC	?=++4B0@FHDB:<+C::CE@HC>?38C9CHAFHGEDHIJCDAFCHGI@FCGBH@@AFEGIIEEHHGH??DFD@>@CA3;?A>A;<<BAC:@CDD######	PG:Z:novoalign	AS:i:3UQ:i:36	NM:i:0	MD:Z:95
          HWI-ST1063_0137:6:1308:10615:65342#0	81	chr1	1019	1	101M	=	765	-354	ATAAATGTCCATAAGTAGACATGAAGCCTGCAGAATTCCAAATAGAATGGACCAGAAAATAAATTCCTCCTGTCACATAATAGTCAAAACACCAAATGCAC	>>;>;5;@;;A;7.))7;7@C=?CA;@DAA=ADC;ECC=DB@IED<DBB?0D99??0499:<9DDC88:2<F9FAFDCA?CCBEE<:C<?DDABDB+??1?	PG:Z:novoalign	AS:i:1UQ:i:14	NM:i:1	MD:Z:14A86	CC:Z:=	CP:i:71209	ZS:Z:R	ZN:i:3	NH:i:3	HI:i:1	IH:i:3
          HWI-ST1063_0137:6:2303:18332:58082#0	419	chr1	1044	1	13S88M	=	1151	210	GAACATAGAAGAAGCCTGCAGAATTCCAAATAGAATGGACCAGAAAATAAATTCCTCCTGTCACATAATAGTCAAAACACCAAATGCACAAAACAAAGAAT	CCCFFFFFHHHHHJJJJJJJGGJJJJJJIGIGJJJIJIIJJJJJJJJIHIEHIDIGIIIIGIGGIJG@GGGGIIIJJHHHHFFFEAACCEEBDDDDACDDC	PG:Z:novoalign	AS:i:105	UQ:i:105	NM:i:0	MD:Z:88	PQ:i:135	SM:i:70	AM:i:70	ZS:Z:R	ZN:i:3	NH:i:3	HI:i:3	IH:i:3
          This is an RNAseq project and we have sufficient number of reads to get a deep sampling. I suspect something is going wrong at this step because down stream I am getting 20% coverage over the genome with this sample...


          I aligned using novoalign, if that is of any consequence.

          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
          30 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 10:19 PM
          0 responses
          32 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 09:21 AM
          0 responses
          28 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-04-2024, 09:00 AM
          0 responses
          53 views
          0 likes
          Last Post seqadmin  
          Working...
          X