Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • onthefly
    Junior Member
    • May 2014
    • 4

    #1

    Bowtie2 exception with rsem

    Hello

    I am using rsem-calculate-expression (with bowtie2) for mapping fastq files (input) to the reference (prepared using rsem-prepare-reference).
    The total number of samples is 6.
    The mapping went fine with the first two samples, but the command got interrupted becuase I accidently switched off my system
    Anyway after resuming the command, I am not able to map any of my remaining 4 samples. For each case the output is:
    Code:
    bowtie2 -q --phred33 --sensitive --dpad 0 --gbar 99999999 --mp 1,1 --np 1 --score-min L,0,-0.1 -I 1 -X 1000 --no-mixed --no-discordant -p 8 -k 200 -x /media/RSEM_Map/reference/ref -1 /media/Fastq/Sample3_R1_001_1.fastq,/media/Fastq/Sample3_R1_002_1.fastq,/media/Fastq/Sample3_R1_003_1.fastq,/media/Fastq/Sample3_R1_004_1.fastq,/media/Fastq/Sample3_R1_005_1.fastq,/media/Fastq/Sample3_R1_006_1.fastq,/media/Fastq/Sample3_R1_007_1.fastq,/media/Fastq/Sample3_R1_008_1.fastq,/media/Fastq/Sample3_R1_009_1.fastq,/media/Fastq/Sample3_R1_010_1.fastq,/media/Fastq/Sample3_R1_011_1.fastq,/media/Fastq/Sample3_R1_012_1.fastq,/media/Fastq/Sample3_R1_013_1.fastq,/media/Fastq/Sample3_R1_014_1.fastq -2 /media/Fastq/Sample3_R2_001_2.fastq,/media/Fastq/Sample3_R2_002_2.fastq,/media/Fastq/Sample3_R2_003_2.fastq,/media/Fastq/Sample3_R2_004_2.fastq,/media/Fastq/Sample3_R2_005_2.fastq,/media/Fastq/Sample3_R2_006_2.fastq,/media/Fastq/Sample3_R2_007_2.fastq,/media/Fastq/Sample3_R2_008_2.fastq,/media/Fastq/Sample3_R2_009_2.fastq,/media/Fastq/Sample3_R2_010_2.fastq,/media/Fastq/Sample3_R2_011_2.fastq,/media/Fastq/Sample3_R2_012_2.fastq,/media/Fastq/Sample3_R2_013_2.fastq,/media/Fastq/Sample3_R2_014_2.fastq | /home//Biotools/sam/samtools view -S -b -o /media/RSEM_Map/Sample3_quals.temp/Sample3_quals.bam -
    Error reading RefRecord 'first' flag
    Error: Encountered internal Bowtie 2 exception (#1)
    Command: /home//Biotools/bowtie2-align --wrapper basic-0 -q --phred33 --sensitive --dpad 0 --gbar 99999999 --mp 1,1 --np 1 --score-min L,0,-0.1 -I 1 -X 1000 --no-mixed --no-discordant -p 8 -k 200 -x /media/RSEM_Map/reference/ref -1 /media/Fastq/Sample3_R1_001_1.fastq,/media/Fastq/Sample3_R1_002_1.fastq,/media/Fastq/Sample3_R1_003_1.fastq,/media/Fastq/Sample3_R1_004_1.fastq,/media/Fastq/Sample3_R1_005_1.fastq,/media/Fastq/Sample3_R1_006_1.fastq,/media/Fastq/Sample3_R1_007_1.fastq,/media/Fastq/Sample3_R1_008_1.fastq,/media/Fastq/Sample3_R1_009_1.fastq,/media/Fastq/Sample3_R1_010_1.fastq,/media/Fastq/Sample3_R1_011_1.fastq,/media/Fastq/Sample3_R1_012_1.fastq,/media/Fastq/Sample3_R1_013_1.fastq,/media/Fastq/Sample3_R1_014_1.fastq -2 /media/Fastq/Sample3_R2_001_2.fastq,/media/Fastq/Sample3_R2_002_2.fastq,/media/Fastq/Sample3_R2_003_2.fastq,/media/Fastq/Sample3_R2_004_2.fastq,/media/Fastq/Sample3_R2_005_2.fastq,/media/Fastq/Sample3_R2_006_2.fastq,/media/Fastq/Sample3_R2_007_2.fastq,/media/Fastq/Sample3_R2_008_2.fastq,/media/Fastq/Sample3_R2_009_2.fastq,/media/Fastq/Sample3_R2_010_2.fastq,/media/Fastq/Sample3_R2_011_2.fastq,/media/Fastq/Sample3_R2_012_2.fastq,/media/Fastq/Sample3_R2_013_2.fastq,/media/Fastq/Sample3_R2_014_2.fastq 
    bowtie2-align exited with value 1
    [samopen] SAM header is present: 31426 sequences.
    [sam_read1] reference 'ID:bowtie2	PN:bowtie2	VN:2.1.0
    ' is recognized as '*'.
    [main_samview] truncated file.
    "bowtie2 -q --phred33 --sensitive --dpad 0 --gbar 99999999 --mp 1,1 --np 1 --score-min L,0,-0.1 -I 1 -X 1000 --no-mixed --no-discordant -p 8 -k 200 -x /media/RSEM_Map/reference/ref -1 /media/Fastq/Sample3_R1_001_1.fastq,/media/Fastq/Sample3_R1_002_1.fastq,/media/Fastq/Sample3_R1_003_1.fastq,/media/Fastq/Sample3_R1_004_1.fastq,/media/Fastq/Sample3_R1_005_1.fastq,/media/Fastq/Sample3_R1_006_1.fastq,/media/Fastq/Sample3_R1_007_1.fastq,/media/Fastq/Sample3_R1_008_1.fastq,/media/Fastq/Sample3_R1_009_1.fastq,/media/Fastq/Sample3_R1_010_1.fastq,/media/Fastq/Sample3_R1_011_1.fastq,/media/Fastq/Sample3_R1_012_1.fastq,/media/Fastq/Sample3_R1_013_1.fastq,/media/Fastq/Sample3_R1_014_1.fastq -2 /media/Fastq/Sample3_R2_001_2.fastq,/media/Fastq/Sample3_R2_002_2.fastq,/media/Fastq/Sample3_R2_003_2.fastq,/media/Fastq/Sample3_R2_004_2.fastq,/media/Fastq/Sample3_R2_005_2.fastq,/media/Fastq/Sample3_R2_006_2.fastq,/media/Fastq/Sample3_R2_007_2.fastq,/media/Fastq/Sample3_R2_008_2.fastq,/media/Fastq/Sample3_R2_009_2.fastq,/media/Fastq/Sample3_R2_010_2.fastq,/media/Fastq/Sample3_R2_011_2.fastq,/media/Fastq/Sample3_R2_012_2.fastq,/media/Fastq/Sample3_R2_013_2.fastq,/media/Fastq/Sample3_R2_014_2.fastq | /home//Biotools/sam/samtools view -S -b -o /media/RSEM_Map/Sample3_quals.temp/Sample3_quals.bam -" failed! Plase check if you provide correct parameters/options for the pipeline!
    Any suggestions/comments on how to resolve this situation are most welcome.

    P.S. Bowtie2 on its own is running fine.

    Thanks
    FlyWorker

Latest Articles

Collapse

  • SEQadmin2
    16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
    by SEQadmin2





    NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification,...
    10-30-2026, 06:46 AM
  • SEQadmin2
    Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
    by SEQadmin2



    Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

    Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
    ...
    10-14-2026, 09:55 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, Yesterday, 07:59 AM
0 responses
16 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-05-2026, 09:25 AM
0 responses
32 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-03-2026, 11:54 AM
0 responses
47 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 10-28-2026, 09:57 AM
0 responses
172 views
0 reactions
Last Post SEQadmin2  
Working...