GSM5374915: D4_BRMKO_REST_ChIPseq; Mus musculus; ChIP-Seq
Cells were lysed, lysates were clarified and incubated with REST antibody(Sigma, 17-10456) for O/N, captured with IgG coated magnetic beads, washed, and ChIPed DNA eluted. Protein-DNA complexes were reverse crosslinked, proteinase digested and ampure bead purified To prepare libraries DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (3 to 5 exo minus) and dATP  and ligated of Illumina adapters. Adapter ligated DNA was PCR amplified with Illumina primers for 14 cycles and library fragments of 200 - 500bp bp were band isolated from an agarose e-gel (invitrogen).  The purified libraries were quantified, and libraries were pooled before sequencing on Illumina NextSeq 500 system