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  • problem with indexing using bfast

    I'm very new to next-gen sequencing and bioinformatics in general. I'm trying to use bfast to align solid4 colourspace reads to a cDNA sequence. I have completed the following steps:

    1) fasta2brg -A 0 -f cDNA.fasta
    2) fasta2brg -A 1 -f cDNA.fasta

    It is when I attemp step 3 that I encounter a problem:

    3) bfast index -A 1 -f cDNA.fa -m 1111111111111111111111 -w 14 -i 1
    bfast index -A 1 -f cDNA.fa -m 111110100111110011111111111 -w 14 -i 2
    bfast index -A 1 -f cDNA.fa -m 10111111011001100011111000111111 -w 14 -i 3
    bfast index -A 1 -f cDNA.fa -m 1111111100101111000001100011111011 -w 14 -i 4
    bfast index -A 1 -f cDNA.fa -m 111111110001111110011111111 -w 14 -i 5
    bfast index -A 1 -f cDNA.fa -m 11111011010011000011000110011111111 -w 14 -i 6
    bfast index -A 1 -f cDNA.fa -m 1111111111110011101111111 -w 14 -i 7
    bfast index -A 1 -f cDNA.fa -m 111011000011111111001111011111 -w 14 -i 8
    bfast index -A 1 -f cDNA.fa -m 1110110001011010011100101111101111 -w 14 -i 9
    bfast index -A 1 -f cDNA.fa -m 111111001000110001011100110001100011111 -w 14 -i 10

    The error message I get is:
    ************************************************************
    Checking input parameters supplied by the user ...
    Validating fastaFileName cDNA.fa.
    Validating tmpDir path ./.
    Input arguments look good!
    ************************************************************
    ************************************************************
    Printing Program Parameters:
    programMode: [ExecuteProgram]
    fastaFileName: cDNA.fa
    space: [Color Space]
    mask: 1111111111111111111111
    depth: 0
    hashWidth: 14
    indexNumber: 1
    repeatMasker: [Not Using]
    startContig: 0
    startPos: 0
    endContig: 2147483647
    endPos: 2147483647
    exonsFileName: [Not Using]
    numThreads: 1
    tmpDir: ./
    timing: [Not Using]
    ************************************************************
    ************************************************************
    Reading in reference genome from cDNA.fa.cs.brg.
    bfast: RGBinary.c:304: RGBinaryReadBinaryHeader: Assertion `rg->contigs[i].contigNameLength > 0' failed.
    /var/spool/torque/mom_priv/jobs/804690.bruce-m.vlsci.unimelb.edu.au.SC: line 10: 4145 Aborted bfast index -A 1 -f cDNA.fa -m 1111111111111111111111 -w 14 -i 1
    ************************************************************

    Is anyone able to suggest what I might be doing wrong? Any suggestions or assistance would be greatly appreciated.

  • #2
    It looks like there is a problem with your input FASTA. Do you have empty lines or empty contigs? Could you post it somewhere for us to take a look?

    Comment


    • #3
      Ah, yes I had a gap in my fasta file between the > and the sample name but I have fixed this up and repeated the first 2 steps and now the third step worked. Thanks

      Comment

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