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  • DNA shearing for NuGEN/Illumina protocol

    Is it absolutely necessary to use a covaris for DNA shearing in the Illumina protocol? I would like to use a NuGen amp followed by Illumina for RNA-seq but do not have a covaris. Has anyone used fragmentase or other enzymatic fragmentation?
    Sridar

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    We use the NuGEN kit a lot and the addition of the shearing step is new to the kit as of the recent version change. It is a very important step however to get good coverage over the transcriptome.

    We have not used an enzymatic fragmentation so I can't comment on how well that would work. I'd be interested to hear your results if you go that way.
    HudsonAlpha Institute for Biotechnology
    http://www.hudsonalpha.org/gsl

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