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  • dmtruong
    Junior Member
    • Dec 2011
    • 1

    Variant Frequency Calculation (non-SNP)?

    Does anyone know of a program to calculate (or count) the number of nucleotides (ATCGindel) at each position of an amplicon alignment (BAM, SAM, or ACE file)?

    It seems that IGV viewer outputs this data in its Coverage Track, and this is also adjustable for percentage variation. However, at this time the data is not exportable, at least, according to IGV help.

    I'm performing a Directed Evolution expt on a 750 nucleotide fragment, and have run 454 titanium reads on the library. Each read has unique nucleotide variation; however, none of the variations exceeds 1% at any position. It seems that most programs are customized for dbSNP calls and call things at much higher percentages.

    I am much less interested in the variant, more so the positions which have the highest variability.

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  • GATTACAT
    Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
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    Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
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  • SEQadmin2
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