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  • Muhammad Afiq
    Junior Member
    • Aug 2015
    • 1
    #1

    16S Library Prep for MiSeq

    Hi all,

    I am a newbie in NGS as well in this community. Currently, I am trying to sequence the human gut microbiota. I have followed the exact Illumina 16S protocol, anyhow the standard protocol PCR produced unspecific band (~1900bp) when analyse it using bioanalyzer. I have tried to modify few thing and try to optimize the PCR program to get rid of the unspecific band/smear but the DNA yield (after purify) is less than the standard protocol. Optimized program also manage to clear off the unspecific band/smear too.

    Any thought on this? I would like to hear some opinion if you guys have any. Thanks
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