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Old 04-09-2013, 11:59 AM   #1
ASL
Junior Member
 
Location: Virginia, USA

Join Date: Apr 2013
Posts: 5
Default NEBNext ChIP-Seq library prep kit vs Illumina TruSeq kit?

Has anyone used the NEBNext ChIP-Seq Library Prep Master Mix Set (or Reagent Set) for Illumina and gotten good results?

I bought this kit on someone else's recommendation, and I'm having my samples sequenced on an Illumina MiSeq in our sequencing core, but the results have been somewhat disappointing: the read number seems to be either lower than expected, or looks very flat (not a lot of the expected peaks). Has anyone else successfully used this kit? Has anyone compared it to the Illumina TruSeq kit?

My source material is PBMCs, the ChIP was done with ENCODE-validated H3K9me3 Ab, and was enriched 22x over the input control. The library itself had a yield of ~45 ng/ul in 15 ul, and was enriched about 400x over the ChIP source material, as validated by real-time PCR, and had an insert size of 274 bp according to the Bioanalyzer chip, so it appeared to be enriched and of the correct size.....

Thanks,
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