GSM1649170: BF2 brain (MeDIP-seq); Heterocephalus glaber; MeDIP-Seq
For MeDIP-seq libraries construction, 5 μg DNA was cut into 100 - 500 bp fragments with a Bioruptor sonicator (Diagenode), followed by end repair, 3’-end addition of dA, and adapter ligation according to Illumina’s protocols. Adaptor-ligated DNA was then immunoprecipitated by a monoclonal anti-methylcytidine antibody. The specificity of the enrichment was confirmed by qPCR using SYBR green mastermix (Applied Biosystems) with primers for positive and negative internal control DNA of non-human samples supplied in the Magnetic Methylated DNA Immunoprecipitation kit (Diagenode). The enriched fragments were purified on ZYMO DNA Clean & Concentrator-5 columns following the manufacturer’s instructions, followed by amplification via 10 adaptor mediated PCR cycles to yield the MeDIP-seq libraries. All MeDIP-seq libraries were subjected to 50 bp paired-end sequencing on an Illumina HiSeq 2000 platform. We analyzed brains from all eight individuals separately, but female ovaries were only large enough for individual analysis in breeders, whereas subordinate ovaries were so small that 3–5 of them had to be pooled for gene expression analyses. The testes of breeding and non-breeding males were not visually different, but breeder status was known because these males had previously sired pups. RNA and DNA were isolated as described in Kim, et al. Nature (2011).
GEO Accession
GSM1649170