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  • gcR
    Member
    • Mar 2017
    • 15

    #16
    You were right, I had been working on that and fixed it alredy. Thanks.

    Now, the featureCounts function says my files are .SAM, altought even checking on bash the class type its .BAM

    It seems like it doesnt open the file and gives 0 count results.

    RESULT:
    Code:
    //========================== featureCounts setting ===========================\\
    ||                                                                            ||
    ||             Input files : 8 SAM files                                      ||
    ||                           S /home/sysadm/tomics/4_GCCAAT_L001_R1_001.f ... ||
    ||                           S /home/sysadm/tomics/8_AGTTCC_L001_R1_001.f ... ||
    ||                           S /home/sysadm/tomics/16_GCCAAT_L002_R1_001. ... ||
    ||                           S /home/sysadm/tomics/20_AGTTCC_L003_R1_001. ... ||
    ||                           S /home/sysadm/tomics/28_GCCAAT_L004_R1_001. ... ||
    ||                           S /home/sysadm/tomics/32_AGTTCC_L004_R1_001. ... ||
    ||                           S /home/sysadm/tomics/40_GCCAAT_L005_R1_001. ... ||
    ||                           S /home/sysadm/tomics/44_AGTTCC_L006_R1_001. ... ||
    ||                                                                            ||
    ||             Output file : ./.Rsubread_featureCounts_pid3679                ||
    ||             Annotations : /home/sysadm/tomics/data3 (GTF)                  ||
    ||                                                                            ||
    ||                 Threads : 1                                                ||
    ||                   Level : feature level                                    ||
    ||              Paired-end : no                                               ||
    ||         Strand specific : no                                               ||
    ||      Multimapping reads : not counted                                      ||
    || Multi-overlapping reads : not counted                                      ||
    ||                                                                            ||
    \\===================== http://subread.sourceforge.net/ ======================//
    
    //================================= Running ==================================\\
    ||                                                                            ||
    || Load annotation file /home/sysadm/tomics/data3 ...                         ||
    ^[[B^[[B||    Features : 1193694                                                      ||
    ||    Meta-features : 58174                                                   ||
    ||    Chromosomes : 25                                                        ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/4_GCCAAT_L001_R1_001.fastq.gz.sub ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/4_GCCAAT_L001_R1_001 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 2380011                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.02 minutes                                             ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/8_AGTTCC_L001_R1_001.fastq.gz.sub ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/8_AGTTCC_L001_R1_001 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 1956680                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.16 minutes                                             ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/16_GCCAAT_L002_R1_001.fastq.gz.su ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/16_GCCAAT_L002_R1_00 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 2287401                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.17 minutes                                             ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/20_AGTTCC_L003_R1_001.fastq.gz.su ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/20_AGTTCC_L003_R1_00 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 2362945                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.17 minutes                                             ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/28_GCCAAT_L004_R1_001.fastq.gz.su ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/28_GCCAAT_L004_R1_00 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 1884255                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.15 minutes                                             ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/32_AGTTCC_L004_R1_001.fastq.gz.su ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/32_AGTTCC_L004_R1_00 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 4205162                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.29 minutes                                             ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/40_GCCAAT_L005_R1_001.fastq.gz.su ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/40_GCCAAT_L005_R1_00 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 1953045                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.14 minutes                                             ||
    ||                                                                            ||
    || Process SAM file /home/sysadm/tomics/44_AGTTCC_L006_R1_001.fastq.gz.su ... ||
    ||    Single-end reads are included.                                          ||
    || WARNING format issue in file '/home/sysadm/tomics/44_AGTTCC_L006_R1_00 ... ||
    ||         The required format is : SAM                                       ||
    ||         The file format is unknown.                                        ||
    || A wrong format may result in wrong results or crash the program.           ||
    || Please refer to the manual for file format options.                        ||
    || If the file is in the correct format, please ignore this message.          ||
    ||                                                                            ||
    ||    Assign reads to features...                                             ||
    ||    Total reads : 2908457                                                   ||
    ||    Successfully assigned reads : 0 (0.0%)                                  ||
    ||    Running time : 0.22 minutes                                             ||
    ||                                                                            ||
    ||                         Read assignment finished.                          ||
    ||                                                                            ||
    \\===================== http://subread.sourceforge.net/ ======================//
    I will keep looking for an answer on the web.

    Thanks all for reading.

    G.
    Beginner @ RNA-Seq, R programming, Linux, Python.-

    Please be patients!

    Comment

    • gcR
      Member
      • Mar 2017
      • 15

      #17
      Found the drama, the BAM files were corrupt, had to DL the dataset again and problem solved.

      Dummy begginers problems, indeed

      Thx for ur time.
      Beginner @ RNA-Seq, R programming, Linux, Python.-

      Please be patients!

      Comment

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