Bisulfite NGS Library – LSU C.virginica Oil Spill MBD Bisulfite DNA Sequencing Submission

Combined the following libraries in equal quantities (17ng each) to create a single, multiplexed sample for sequencing (LSU_Oil_01):

  • HB2 – 1 (ATCACG)
  • HB16 – 3 (TTAGGC)
  • HB30 – 4 (TGACCA)
  • NB3 – 5 (ACAGTG)
  • NB6 – 6 (GCCAAT)
  • NB11 – 7 (CAGATC)

Quantified pooled libraries using the Quant-iT dsDNA BR Kit (Invitrogen) with a FLx800 plate reader (BioTek). Used 1μL of the pooled sample, run in duplicate. Used 1uL of standards, run in duplicate.

Results:

pooled libraries = 6.575ng/μL

Will submit to University of Oregon Genomics Core Facility for 100bp, single end Illumina HiSeq2500 sequencing. They need 10nM of sample. For a library with average size range of 300-400bp, this requires a sample volume of 20uL with a concentration of 2.28ng/μL in a solution of 0.1% Tween20 in Buffer EB (Qiagen).

Combined 6.94μL of pooled libraries with 13.06 of 0.1% Tween20/EB solution.

Submitted sample LSU_Oil_01 to University of Oregon Genomics Core Facility via O/N FedEx on dry ice. Sample was assigned order # 2112.

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