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  • DNA Tru Seq with cDNA

    I am aware that there are kits expressly for mRNA Tru Seq, but we are attempting to use a DNA Tru Seq kit starting with cDNA. Does anyone have experience with this or know off hand the minimum starting concentration of gDNA that the kit requires?

  • #2
    Well, 1 ug is the amount specified in the protocol. But I guess it will work fine down to 100 ng or so without modifying anything. Below that you probably want to start diluting your adapters to keep the adapter-dimer concentration low.

    Also, check out this thread. Using the numbers that Ethanol gives would lead one to believe that the TruSeq RNA kit expects about 25 ng of ds cDNA to be generated.

    Anyway, within reason, you could just dilute your TruSeq DNA adapters down by what ever factor your input DNA is below the 1 ug specified in the protocol. My guess is that will work down to 10 ng or so. Maybe lower...

    --
    Phillip

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