--- author: Sam White toc-title: Contents toc-depth: 5 toc-location: left date: 2015-01-28 04:03:20+00:00 layout: post slug: bisuflite-ngs-library-prep-c-gigas-larvae-oa-bisulfite-dna-continued-from-yesterday title: Bisuflite NGS Library Prep - C.gigas larvae OA bisulfite DNA (continued from yesterday) categories: - 2015 - Crassostrea gigas larvae OA (2011) bisulfite sequencing tags: - barcodes - BS-seq - Crassostrea gigas - library prep - Pacific oyster --- Continued Illumina library prep of bisulfite-treated DNA samples (400ppm and 1000ppm; from [20150114)(2015/01/14/dna-bisulfite-conversion-c-gigas-larvae-oa-sheared-dna.html)  with [Methylamp DNA Modification Kit (Epigentek)](https://github.com/sr320/LabDocs/blob/master/protocols/Commercial_Protocols/Epigentek_PostBisulfiteIlluminaLibraryPrep_P-1055.pdf). Performed bead clean up immediately after End Repair. PCR cycles: 14 No other changes were made to the manufacturer's protocol. Epigentek Barcode Indices assigned, per their recommendations for using two libraries for multiplexing: 400ppm - barcode #6 - GCCAAT 1000ppm - barcode #12 - CTTGTA The two libraries were stored @ -20C and will be quantified tomorrow.