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  • 454 assembly using consed

    Has anyone assembled 454 data with consed package version 19? I'm having some issues with reading of the .sff files and wondering if anyone has completed an assembly of 454 read data (not using Roche software produced .ace files). I'm using "add454Reads.perl reference.ace sff.fof reference.fa", where the fof specifies the sff files to assembly, but although the script runs, I get an error "doesn't existile /shared/BNFinal/mapping/consed/sff_dir/FPDLD6P02.sff", and the 454 reads are not brought into the assembly; it basically assembles with only the reference sequence. I also tried specifying just the names of reads that I want to bring in to the assembly; these exist in the sff files in the corresponding sff_dir directory to the edit_dir I am running the assembly from. Someone mentioned needing to update phrap, which I will look into, but any other thoughts on this (I'm sure I'm missing something obvious)?
    Thanks,
    Liz

  • #2
    I have successfully assembled 454 data with consed version 19? sff.fof contains the pathway of *.sff file. do you have the permission to access the sff file?

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    • #3
      Thanks for the response. Could you give an example of the contents of the sff.fof file? I checked permissions, but will check again!
      Liz

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      • #4
        the content is :
        ../sff_dir/FPDLD6P02.sff
        sff.fof, add454Reads.perl, reference.ace reference.fa are all in edit_dir.
        if your reads are too much, you need use cross_match.manyreads

        good luck

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        • #5
          Well still not sure what I'm doing wrong. The permissions for the files and directories are fine. The sff.fof file itself seems to be fine; the contents are being read. The sff file looks normal (can load it into other apps etc.), and the path is correct..... Thanks!

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          • #6
            add454Reads.perl command is "consed". Have you change consed_linux64bit to consed?
            Last edited by bio-x; 06-05-2009, 05:55 AM.

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            • #7
              Dear sir!, bio-X

              please tell me how to configure the consed 19 for the assembly of 454.sff file , as it was 6 month i am trying to assemble but cannot get the success .

              sir please tell the procedure how to configure the phred ,phrap and consed to assemble, .sff files

              and the necessary version needed for the assembley of the .sff files..please sir it was great help for me. as my boss is annoyed on me very much

              i shall be very thankful to you

              waiting for your kind reply please mail me
              [email protected]

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              • #8
                Originally posted by ajaybioinfo View Post
                Dear sir!, bio-X
                please tell me how to configure the consed 19 for the assembly of 454.sff file , as it was 6 month i am trying to assemble but cannot get the success .
                What have you tried? What problems occured? Have you carefully read the documentation supplied with the package (http://bozeman.mbt.washington.edu/co...EADME.19.0.txt)?

                sir please tell the procedure how to configure the phred ,phrap and consed to assemble, .sff files
                Again, have you read the docs?


                and the necessary version needed for the assembley of the .sff files..
                You should use at least consed v19.xx and cross_match version 1.080806 or higher.

                Assembling 454 data with consed usually means "reference guided" assembly, not de-novo. You should keep that in mind.

                cheers,
                Sven

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