title: GSM2827680: control C4; Danio rerio; Bisulfite-Seq
library_construction_protocol: Genomic DNA from each pooled larvae sample was extracted using DNeasy Blood & Tissue Kit (Qiagen) following the manufacturer's protocol. Reduced representation bisulfite sequencing libraries were prepared based on published protocols. Briefly, 12 RRBS libraries were prepared, 4 libraries per condition, each library containing a pool of 24 larvae. Genomic DNA was digested with MspI (New England Biolabs, Ipswich, MA) followed by end repair, addition of 3′ A overhangs and addition of methylated adaptors (Illumina, San Diego, CA) to the digested fragments. Following adaptor ligation, DNA fragments ranging from 40–220 bp (pre-ligation size) were cut from a 3% (w/v) NuSieve GTG agarose gel (Lonza, Basel, Switzerland) and subsequently bisulfite converted using the EZ DNA methylation kit (Zymo Research, Irvine, CA) with an extended incubation time of 18–20 h. Bisulfite converted libraries were amplified by PCR reactions and sequenced (100-bp single ended reads) by New Zealand Genomics Limited (University of Otago, New Zealand) on an Illumina HiSeq2000 sequencer.
attributes.tag: GEO Accession
attributes.value: GSM2827680