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Old 08-01-2015, 09:09 PM   #1
Muhammad Afiq
Junior Member
 
Location: Singapore

Join Date: Aug 2015
Posts: 1
Default 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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