GSM1411206: RPB2_PolII_ZT02_R1; Mus musculus; ChIP-Seq
A total of 1 ml of each chromatin suspension (containing about 60 mg of DNA) was incubated with 10 mg of anti-RPB2, 1.5 mg of anti-H3K36me3, or 1.5 mg of anti-H3K4me3 in buffer A (20 mM Tris/HCl (pH 7.5), 150 mM NaCl, 2 mM EDTA) overnight at 4uC on a rotating wheel. Ten ml of protein A bead suspension (25% slurry in buffer A), pre-blocked with 10 mg/ml of salmon sperm DNA and BSA at 4uC overnight, was then added and the incubation was continued for 1 h at room temperature on a rotating wheel. The beads were then washed with dialysis buffer and ChIP wash buffer. Protein–DNA complexes were eluted from the beads, de-cross-linked, and treated with RNase A and, subsequently, with proteinase K. A total of 10–12 ng DNA were used for the preparation of the library. Libraries for ultra-high throughput sequencing were prepared with the ChIP-Seq DNA sample kit (Illumina) as recommended by the manufacturer.
GEO Accession
GSM1411206