title: GSM5365935: HA36_ATF7IP-KO_ES; Mus musculus; Bisulfite-Seq
library_construction_protocol: Genomic DNA extraction, followed by phenol/chloroform extraction and ethanol precipitation. RNaseA treatment was performed for 30 min at 37C Gemomic DNA was converted using the Qiagen Epitect bisulphite conversion kit. After conversion, two separate PCR amplifications per target were performed at 50 ˚C and 58 ˚C annealing temperature with the PhusionU polymerase (Thermo Scientific) for 45 cycles of 1 min 95 ˚C denaturation, 1 min for annealing, and 1 min 72 ˚C for extension. PCR products were run on a 1.2 % agarose gel, extracted, quantified and pooled at equimolar ratios. Library preparation was done with 10 ng of input material using the NEBNext Ultra II kit protocol and cleaned up using Ampure XP beads (Beckman Coulter). targeted bisuphite PCR- seq