GSM2064389: RNAseq_HG-mESCs; Mus musculus; RNA-Seq
ChIP-enriched DNA was obtaind by using ChIP-IT Express Enzymatic (Active Motif, 53009) and S40Gc specific antibody (20B2). Total RNA was isolated from cells using Direct-zol RNA Kit (Zymo Research, R2052) according to the manufacturer’s instructions. The ends of 50 ng of ChIP-enriched DNA and Input DNA were repaired using T4 DNA polymerase (New England Biolabs, M0203) and phosphorylated with T4 polynucleotide kinase (New England Biolabs, M0201). A single adenine base was added to the 3 prime-end with Klenow fragments (New England Biolabs, M2012). TruSeq DNA adapters (Illumina, FC-121-2001/-2002) were ligated to the fragments with T4 DNA ligase (New England Biolabs, M2200). Ligation products between 200 and 600 bp were purified on AMpure XP beads (Beckman Coulter, A63880) to remove unligated adapters and subjected to 14 cycles of PCR amplification by KAPA Library Amplification Kit (KAPA BIOSYSTEMS, KK2611). RNA-seq libraries were constructed using TruSeq Strand mRNA LT Sample Prep Kit (Illumina) according to the manufacturer’s instructions. Completed libraries were quantified using an Agilent 2000 BioAnalyzer system.
GEO Accession
GSM2064389