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  • single-cell cDNA left 90 min at 95°C

    Briefly, cDNA from single neurons prepared using SMART-Seq4 kit spent 90 minutes at 95°C due to human error. Is the cDNA to be discarded or can it still be used for library preparation with Nextera XT and sent to sequencing?

    Thanks

  • #2
    Prolonged high heat exposure definitely introduces a lot of damages to DNA. I would expect it to be pretty fragmented by now. I also assume the Nextera transpose would require dsDNA - thus, you would need to add at least another PCR cycle. Perhaps you could use this S1 kit instead?: https://swiftbiosci.com/products/acc...-library-kits/

    If the sample can be reproduced, best dump it.
    Last edited by luc; 07-13-2018, 05:55 PM.

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