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  • gene copy number -k parameter

    hi,
    I have 3 questions on the same problem:
    I'm aligning reference DNAseq (illumina) to itself. I plot the gene coverage histogram and try to calculate the gene copy number assuming the peak represents single copy.
    1) how does it happen that i get genes with 30 copies even tho i only allow a single alignment per read to be reported (-k 1)? one explanation is that the genome annotation is not correct, and that there are more copies for some genes than is actually annotated. Is there any other explanation?
    2) Using -k30 parameter gives a gene coverage histogram of Poisson distribution with a very long right tail. Using the -k1 parameter gives a histogram with 2 peaks. How can I explain the 2 peaks (a small one followed by the big one, which doesn't fit the fact that the majority of genes have one copy, and a smaller number have 2, etc.)
    3) I realize the gene coverage will not be the same in the 2 runs, but why does the order of genes with high-low coverage differ as well?
    thanks,

  • #2
    which software did you use for the coverage histogram?

    I am working with DNA copy number and am brand new to this type of analysis.

    I have a coverage file generated by IGV. how did you generate your coverage data?

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