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  • spikey_hedgehog
    Junior Member
    • Feb 2012
    • 5

    digital PCR & ChIP-seq (internal control?)

    Hi guys,

    I am very new to making my own libraries and I am interested in using digital PCR to QC my ChIP-seq libraries. So I would have the universal primers spanning the adaptors and run the diluted libraries on the chip.

    Is it necessary to have an internal control for this? Other people have used RNAse P but that's only if I know RNAse P will be enriched in my library..right?

    How do people choose their internal control (if they have one)?

    Thanks heaps! Look forward to your answers!

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  • GATTACAT
    Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
    by GATTACAT
    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
    Nine Things a Sample Prep Scientist Thinks About Before Sequencing
    by SEQadmin2


    I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

    Here are nine questions we think about, in roughly the order they matter, before...
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