title: GSM3374746: C5_methylation; Bos taurus; Bisulfite-Seq
library_construction_protocol: Total DNA was extracted from liver and mammary gland samples for reduced representation bisulfite sequencing analysis. Extraction, library construction, bisulfite treatment and sequencing were performed by Epigentek. DNA was isolated from 50 mg of tissue with the FitAmpTM General Tissue Section DNA Isolation Kit. DNA was digested with MspI restriction enzyme for 2 hrs at 37°C followed by digestion with TaqαI restriction endonuclease at 65°C. DNA fragments < 300 bp were selected and used for bisulfite treatment with the Methylamp DNA Conversion Kit. Standard lllumina sequencing adaptors were ligated onto the bisulfite treated DNA fragments using a random probing method. Library amplification was achieved using indexed primers and analyzed on a Bioanalyzer and with a KAPA Library Quantification Kit according to the manufacturer's protocol. Purified DNA (15 nM of sample) libraries were sequenced using the Illumina HiSeq 4000 system generating 50 bp single-end reads.
attributes.tag: GEO Accession
attributes.value: GSM3374746