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  • sehrrot
    Member
    • Jul 2010
    • 58

    Library prep for 50ng input with TruSeq DNA- or RNAseq adapters?

    Dear all,

    I am going to process library preparation with 50 ng DNA. I am just concerning ratio of adaptersNA input. Thus, I am considering to use adapters from TruSeq RNAseq kit, which has lower molarity adapters. Has anyone done library prep with lower DNA input?
  • pmiguel
    Senior Member
    • Aug 2008
    • 2328

    #2
    Yes or you could just dilute your DNA TruSeq adapters 20x.

    We have gone below 1 ug. Possibly to 100 ng, with no modifications to the protocol. But we generally size select prior to enrichment PCR. So adapter dimers are not that much of an issue.

    --
    Phillip

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      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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