GSM3721759: D_ES_F_rep2; Bos taurus; Bisulfite-Seq
The WGBS libraries were constructed following the standard protocol. Briefly, genomic DNA with 0.5%unmethylated lambda DNA (Fermentas) spike-in was first sonicated into 200–300-bp fragments using Covaris S2 system. After being concentrated, the sheared DNA was end-repaired, dA-tailed and ligated with pre-methylated TruSeq DNA adapters (illumina). Bisulfite conversion was conducted with Methyl Code Bisulfite Kit (Invitrogen), after Bisulfite conversion, the converted templates were PCR amplified and quantified using Qubit ds DNA high sensitivity dye (Invitrogen). For RNA-seq library, Total RNA was extracted from 4*10^6 cells using RNeasy Mini Kit according to the manufacturer's instructions. The mRNA was isolated from 2 μg of total RNA using the NEB Next Poly(A) mRNA Magnetic Isolation Module. Subsequently, isolated mRNA was used for RNA library construction using NEB Next ultra RNA library prep kit for Illumina. The final quality-insured libraries were sequenced on HiSeq2000 sequencer.
GEO Accession
GSM3721759