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  • Daisy-Fu
    Junior Member
    • May 2011
    • 4

    #1

    How to dig out single particular gene from assembled contigs

    Sorry if this question has been asked before. I just did a RNA-seq of a plant parasitic nematode species, and assembled 50,000 contigs by Trinity.
    I would like to dig out a full-length sequence of endoglucanase gene, and do some protein structure prediction like that. Partial sequence of this gene was available through previous degenerate PCR. Does anybody know a easy and simple way to do it?
    Thanks a lot.
  • westerman
    Rick Westerman
    • Jun 2008
    • 1104

    #2
    Blast of your Trinity contigs against the partial sequence? Should be fast enough and sensitive enough to pick up the contig(s). Or am I missing something obvious here?

    Comment

    • Daisy-Fu
      Junior Member
      • May 2011
      • 4

      #3
      Originally posted by westerman View Post
      Blast of your Trinity contigs against the partial sequence? Should be fast enough and sensitive enough to pick up the contig(s). Or am I missing something obvious here?
      Hey, Westerman in Purdue, thank you for you replying.
      Are you talking about build a local db or www-blast something like that?

      Comment

      • westerman
        Rick Westerman
        • Jun 2008
        • 1104

        #4
        Local db. Using the older version of blast (although blast+ is similar):

        1) formatdb -i Trinity.fasta -p F -o T
        2) blastall -p blastn -d Trinity.fasta -i myfile.fa -o output.blastn
        3) manually look at output.blastn to figure out the id of the matching contig
        4) fastacmd -d Trinity.fasta -s myid > found.fa

        Comment

        • cewells
          Junior Member
          • Jul 2011
          • 3

          #5
          Thanks westerman -- we modified for the new blast+ as below --

          1) makeblastdb -in Trinity.fasta -dbtype nucl -parse_seqids -out TrinityBlast -title "All contigs"
          2) blastn -query myfile.fa -db TrinityBlast -out myfile.out
          3) same
          4) blastdbcmd -db TrinityBlast -entry myid > found.fa

          Comment

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