title: GSM2044558: TFH92_WGBS; Macaca mulatta; Bisulfite-Seq
library_construction_protocol: genomic DNA was extracted following the Qiagen DNeasy Blood & Tissue Kit  Paired-end BS-seq libraries for Illumina sequencing were constructed according to Lister et al.(2009) with minor modifications. In brief, 300 - 500 ng of genomic DNA was fragmented by sonication, end repaired and ligated to custom-synthesized methylated adapters (Eurofins MWG Operon, Huntsville, AL) according to manufacturer’s (Illumina, San Diego, CA) instructions for gDNA library construction. Adaptor-ligated libraries were subjected to two successive treatments of sodium bisulfite conversion using the EpiTect Bisulfite kit (Qiagen, Valencia, CA) as outlined in the manufacturer’s instructions. Five to ten ng of bisulfite-converted libraries was PCR amplified with the following condition: 2.5 U of ExTaq DNA polymerase (Takara), 5 μl of 10X Extaq reaction buffer, 25 μM dNTPs, 1 μl Primer 1.1, 1 μl Primer 2.1 (50 μl final). The thermocyling was as follows: 95°C 3 min, then 10 cycles of 95°C 30 sec, 65°C 30 sec and 72°C 60 sec. The enriched libraries were purified twice with SPRI method using AMPure beads (Beckman Coulter, Inc. Brea CA).
attributes.tag: GEO Accession
attributes.value: GSM2044558