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  • strob
    Member
    • Nov 2008
    • 84

    #1

    filter high quality solexa reads

    Dear all,

    I just received data from illumina (100nt reads).
    The file is named: mRNAseq_7_sequence.txt
    and it looks like this:

    HWUSI-EAS1999_9996:7:1:2:618#0/1:NATGTTTTTTTTTTTCAAGAACGAAAGTTNGGGGCTCGAAGACGATCAGATACCGAGAAAAAAAAAAAATCGTATGCCGTCTTCTGCTTGAAAAAAAAAAA:FNNNMWWTTYeeeeeeeeeeeeeeeRRTTEVVUUUeeeeeeeeeeeeeeeeeeeeeeee\eeeeeee[ee^\\[[\^\\\\^ZY[eee[eeeeeeBBBBB
    HWUSI-EAS1999_9996:7:1:2:1071#0/1:GACGACTTCTCCGGGGGGGAAATGATAAGNTTCAGTGGACTTCCCCCCCCCCCGCGGGCAGCGAAAAAAAAAAAAAAACCGAGATCGGAAGAGCTCGTATG:EHHHHMMJJJeeeeeeeeee\eeeeKKJJEPNNPP\^^Y\^\^^Yeeeee\[\\[ee[e[\\^[\eeeeeBBBBBBBBBBBBBBBBBBBBBBBBBBBBBB
    HWUSI-EAS1999_9996:7:1:2:264#0/1:NGGCCATCAGTAGGGTAAAAAAAAAAAAANTCACGACGGTCTAAACCCAGCTTTTTTTTTTGAAGAGCTCGTATGCCCCCCCCCCCTTGAAAAAAAAAAA:FTVVV^^Y^^eeeeeeeeeeeeeee^^^ZFRRVROeeeeeeeeeeeeeeeeee[eeeeeeeeeeeeeeeeee\eeee[eeeeeee\e[[e\[^BBBBBBB


    I have been searching all over the net, but there is too much info, so here is my question: how can I filter for high quality reads? I want to filter for reads which have or an average quality above a certain threshold or have a certain number of bases below a certain threshold. Is there a way to do this?

    Next I want to convert this to 1. solexa fastq ; 2. normal fastq; ...

    As I want to use this data in GALAXY, I should also be able to upload it there.

    Any help is more than welcome

    Steven
  • svl
    Member
    • Sep 2009
    • 43

    #2
    Originally posted by strob View Post
    Next I want to convert this to 1. solexa fastq ; 2. normal fastq
    Normally the _sequence.txt files are in fastq format...
    -----
    @GAII:6:1:50:677#0/1
    ATGCTT etc
    +GAII:6:1:50:677#0/1
    Z]^_^^ etc
    -----
    ...so why didn't they just give you those?

    Anyway, to get fastq back, you would have to split every line in your file into four parts and print back
    @header
    sequence
    +header
    quality-values

    And when you have fastq, you can indeed use galaxy to trim your reads based on quality

    -svl
    Last edited by svl; 12-08-2009, 06:23 AM.

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