Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • Hyzteria
    Junior Member
    • Oct 2018
    • 1

    #1

    BS-seeker2 alignment error (bowtie2)

    Hi everyone ,

    I'm trying to align some of ours RRBS data on human genome hg38 with BS-seeker2.
    Indexing went good without any error except the warning :
    Code:
    Warning: Encountered reference sequence with only gaps
    but I know why.

    I get errors in the alignement step about some file or directory not found, as you can see :

    Code:
    -bash-4.1$ ./bs_seeker2-align.py -i /YYYYYYY/NNRD204.fastq.gz --aligner=bowtie2 -o PostCT_6m_bs-seeker2.bam -g ../../sequence/hg38.fa -r -L 20 -U 250 -t Y
    
         BS-Seeker2 v2.1.5 - Dec. 21, 2017
    
    [2018-10-10 11:24:42] Mode: Bowtie2, local alignment
    [2018-10-10 11:24:42] Filter for tag XS: #(mCH)/#(all CH)>50.00% and #(mCH)>5
    [2018-10-10 11:24:42] Temporary directory: /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M
    [2018-10-10 11:24:42] Reduced Representation Bisulfite Sequencing: True
    [2018-10-10 11:24:42] Single end
    [2018-10-10 11:24:42] Aligner command: None/bowtie2 --local --quiet -p 2 -D 50 --norc --sam-nohead  -k 2 -x %(reference_genome)s -f -U %(input_file)s -S %(output_file)s
    [2018-10-10 11:24:42] ----------------------------------------------
    [2018-10-10 11:24:42] Read filename: /YYYYYYYY/NNRD204.fastq.gz
    [2018-10-10 11:24:42] The first base (for mapping): 1
    [2018-10-10 11:24:42] The  last base (for mapping): 200
    [2018-10-10 11:24:42] Path for short reads aligner: None/bowtie2 --local --quiet -p 2 -D 50 --norc --sam-nohead  -k 2 -x %(reference_genome)s -f -U %(input_file)s -S %(output_file)s
    
    [2018-10-10 11:24:42] Reference genome library path: /XXXXXX/BS-seeker2/bs_utils/reference_genomes/hg38.fa_rrbs_20_250_bowtie2
    [2018-10-10 11:24:42] Un-directional library
    [2018-10-10 11:24:42] Number of mismatches allowed: 4
    [2018-10-10 11:24:42] --------------------------------
    [2018-10-10 11:24:42] Start reading and trimming the input sequences
    [2018-10-10 11:24:55] Processing read file: /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/NNRD204.fastq.gz-s-1
    [2018-10-10 11:25:09] Processing input is done
    [2018-10-10 11:25:09] Start mapping
    [2018-10-10 11:25:09] Starting commands:
    [2018-10-10 11:25:09] Launched: None/bowtie2 --local --quiet -p 2 -D 50 --norc --sam-nohead  -k 2 -x /XXXXXX/BS-seeker2/bs_utils/reference_genomes/hg38.fa_rrbs_20_250_bowtie2/W_C2T -f -U /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/Trimmed_C2T.fa.tmp-9757942 -S /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/W_C2T_m4.mapping.tmp-9757942
    [2018-10-10 11:25:09] Launched: None/bowtie2 --local --quiet -p 2 -D 50 --norc --sam-nohead  -k 2 -x /XXXXXX/BS-seeker2/bs_utils/reference_genomes/hg38.fa_rrbs_20_250_bowtie2/C_C2T -f -U /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/Trimmed_C2T.fa.tmp-9757942 -S /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/C_C2T_m4.mapping.tmp-9757942
    [2018-10-10 11:25:09] Launched: None/bowtie2 --local --quiet -p 2 -D 50 --norc --sam-nohead  -k 2 -x /XXXXXXX/BS-seeker2/bs_utils/reference_genomes/hg38.fa_rrbs_20_250_bowtie2/W_G2A -f -U /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/Trimmed_G2A.fa.tmp-9757942 -S /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/W_G2A_m4.mapping.tmp-9757942
    [2018-10-10 11:25:09] Launched: None/bowtie2 --local --quiet -p 2 -D 50 --norc --sam-nohead  -k 2 -x /XXXXXXX/BS-seeker2/bs_utils/reference_genomes/hg38.fa_rrbs_20_250_bowtie2/C_G2A -f -U /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/Trimmed_G2A.fa.tmp-9757942 -S /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/C_G2A_m4.mapping.tmp-9757942
    
    Traceback (most recent call last):
      File "./bs_seeker2-align.py", line 367, in <module>
        options.cut_format
      File "/XXXXXXX/BS-seeker2/bs_align/bs_rrbs.py", line 682, in bs_rrbs
        'output_file' : CG2A} ])
      File "/XXXXXXX/BS-seeker2/bs_utils/utils.py", line 332, in run_in_parallel
        for i, proc in enumerate([subprocess.Popen(args = shlex.split(cmd), stdout = stdout) for cmd, stdout in commands]):
      File "/local/miniconda3/envs/python-2.7.15/lib/python2.7/subprocess.py", line 394, in __init__
        errread, errwrite)
      File "/local/miniconda3/envs/python-2.7.15/lib/python2.7/subprocess.py", line 1047, in _execute_child
        raise child_exception
    [B]OSError: [Errno 2] No such file or directory[/B]

    So I tried to launch bowtie2 alignements with the commands just preciding the error (without the --quiet flag) and it went good, no error to declare :
    Code:
    bowtie2 --local -p 2 -D 50 --norc --sam-nohead  -k 2 -x /XXXXXX/BS-seeker2/bs_utils/reference_genomes/hg38.fa_rrbs_20_250_bowtie2/W_C2T -f -U /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/Trimmed_C2T.fa.tmp-9757942 -S /tmp/bs_seeker2_PostCT_6m_bs-seeker2.bam_-bowtie2-local-TMP-FAvs7M/W_C2T_m4.mapping.tmp-9757942
    I don't get why it raises me this error. I tried on local and on a distant cluster .
    I would be very grateful if someone could help me !

    Have a nice day !

Latest Articles

Collapse

  • kinpict.com
    Reply to 16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
    by kinpict.com
    The part about this topic feels surprisingly grounded, and the details on the page is the bit that made me slow down. I’d keep <a href="https://kinpict.com/">ai family photo generator</a> bookmarked for later.
    Today, 06:37 PM
  • 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

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, 11-12-2026, 07:59 AM
0 responses
26 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-05-2026, 09:25 AM
0 responses
35 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-03-2026, 11:54 AM
2 responses
55 views
0 reactions
Last Post kinpict.com  
Started by SEQadmin2, 10-28-2026, 09:57 AM
0 responses
175 views
0 reactions
Last Post SEQadmin2  
Working...