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overrepresented reads with a given length



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  • overrepresented reads with a given length


    we sequence human DNA with 454 Junior, single-read. Runs are ok (number and quality of reads).
    After trimming (quality>20 or 25 according to tests) and removing adapters (15nt), the reads distribution according to the length is quite a Gauss curve except a peak at the max reads number for a given size. This peak describe reads with the same length for most passed runs (499nt if qual>20, 483nt if qual>25). We don't observe this peak before trimming and these reads (as runs) match with our interest region (no contamination).
    Ours questions:
    - why is there a peak and not something like a Gauss curve?
    - why always the same length (after trimming) regardless runs?
    Can we say that the polymerase enzyme and reagents are really as reproducible as possible that, at the cycle n=X, their quality/efficiency falls?
    quite a planned obsolescence of biological (and commercial) products

    Thanks for your help!

    > problem solved by biologist (error with the sonication step...)
    Last edited by ebe; 06-07-2015, 11:32 PM. Reason: solved problem

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