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  • Kevin_YY
    Junior Member
    • Sep 2010
    • 2

    Need reverse reads or not?

    Dear All,

    When we assemble sequences by using program such as MIRA or Velvet, do we need to make sure that all the reverse reads were turned into the forward one? Or just put all the forward and reverse reads in one fasta file.

    Thx
  • maubp
    Peter (Biopython etc)
    • Jul 2009
    • 1544

    #2
    What kind of paired end data are you starting with?

    For velvet, you need one file for paired reads where they are interleaved (forward read then reverse read), and a separate file for any single reads.

    For MIRA, you put all the paired reads in the same input file (either a FASTA + matching QUAL file, or a FASTQ file). The reads must be named so as to identify the pairs, and yes, the reverse reads should be inwards as though done with Sanger end sequencing.

    Basically read the MIRA or Velvet documentation

    Comment

    • Torst
      Senior Member
      • Apr 2008
      • 275

      #3
      Originally posted by maubp View Post
      For velvet, you need one file for paired reads where they are interleaved (forward read then reverse read), and a separate file for any single reads.
      Actually Velvet does not care what direction they face, as long as they are opposite... for example, if you feed it (<= =>) mate pairs, it can estimate (or you can supply) an insert size of -3000 bp (negative 3000) ... but I wouldn't trust the code completely!

      Originally posted by maubp View Post
      Basically read the MIRA or Velvet documentation
      If people start RTFM-ing, then SeqAnswers will die! ;-)

      Comment

      • anyone1985
        Member
        • Mar 2009
        • 68

        #4
        you mean velvet can reverse the reads itself, even if it is mate pairs. however, in the mailing list, more than one time, they have mentioned to reverse both of the two reads to assemble the mate pairs

        Originally posted by Torst View Post
        Actually Velvet does not care what direction they face, as long as they are opposite... for example, if you feed it (<= =>) mate pairs, it can estimate (or you can supply) an insert size of -3000 bp (negative 3000) ... but I wouldn't trust the code completely!



        If people start RTFM-ing, then SeqAnswers will die! ;-)

        Comment

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