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  • Problem generating consensus files

    Hello all,

    I have been having trouble generating consensus files from sorted BAM files.

    I have tried both the mpileup command-

    samtools mpileup -uf ./genomes/NC_013316.fna 2245.sorted.bam | ../samtools-0.1.16/bcftools/bcftools view -cg - | ../samtools-0.1.16/bcftools/vcfutils.pl vcf2fq > 2245cns.fq

    and the depreciated pileup command -

    samtools pileup -cf ./genomes/NC_013316.fna 2245.sorted.bam | ../samtools-0.1.16/misc/samtools.pl pileup2fq -D150 > 2245pcns.fastq

    Both commands run correctly, however when I look at the fastq file it truncates at line 69857 and the rest of the files is filled with this character ( ~~~~) and random letters, as shown below -

    TCCATAAATTTAGTTATTCTGTAAAATTTATATATATTTTTGCTTTTCTGTTAATAACTT
    TGTGGAAAATATTAATTATTCTGTTCGGAGGTTTATatt
    +
    ?BBEEEEHNNNNNZZ`ccffiiloru~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
    ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
    ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~

    Can anyone please explain what may be going on?
    I am very new to seq analysis and can't work it out!

    Emma

  • #2
    please ignore the last post!

    PLease ignore the last post, I have just realised that I have not converted my file to fasta, therefore I still have the quality called in the fastq file!

    Comment


    • #3
      samtools to convert Bam file to consensus sequence

      Hi, all:
      I tried to covert Bam file to consensus sequence as follow command:

      Samtools pileup -cf primatecom-sorted.bam samtools.pl pileup2fq -D100 > primate.fa

      error as follows:
      the Pileup command has been removed. please use mpileup instead.

      according to EmmBB, I tried mpileup. However, I didn't find samtools-0.1.18/bcftools/bcftools.

      Could you help me for this matter?

      Thank you very much,

      Runhua

      Comment

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