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  • graham90978
    Junior Member
    • Oct 2010
    • 6

    #1

    Problem with de novo paired-end data

    Hi guys I have a problem that I cannot seem to get around regarding Paired end vs Single end data. I have a data set of ~1000 genes with an average length of 900bp per gene.
    I wish to compare the various read lengths and their affect on de novo assembly of multiple genes in bot SE and PE data.
    I have simulated various length data sets using the ART NGS simulator. The command that I used for the PE data was :

    ./art_illumina --paired -i genes.fa -l 85 -f 200 -m 900 -s 150 -o outfile.fa
    where reads of length 85 will be generated with fold coverage of 200 and mean DNA fragment size of 900 with a standard deviation of 150.

    I have used SOAPdenovo to assemble all data however, more of the initial genes are recovered using SE data. I would have expected PE data to produce better results, it is quite poor in comparison to SE.

    I think I may be doing something wrong in the simulation as the final assembled contigs only ever get to a max size of about 400bp for PE.

    Does anyone know what the problem might be?
    Thank you!
  • mruizm
    Member
    • Apr 2013
    • 22

    #2
    Illumina MiSeq sequencing

    Hi everybody, i'm confussed about how can i verify if the information obtained from a MiSeq Illumina sequencing of a transcriptome is robust or not.
    I have a sequencing of three tissues obtaining a total of 29.603.685 reads of 151 bp lenght and an assembly of the reads, obtaining a total of 64.924 transcripts with a total of 53.409.526 bp. I need to know if the number of reads used are enough to verify if the information is robust or not.

    Could you help me with that? Please help me with a reference too!

    Best regards!

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