GSM5841357: sg4841 Paf1KD Ints11 ChIP-seq; Mus musculus; ChIP-Seq
Briefly, cells were crosslinked at room temperature for 10 min with 1% formaldehyde,  washed twice with PBS and sonicated in lysis buffer (50 mM HEPES/KOH pH 7.5, 140  mM NaCl, 0.1% Na-Deoxycholate, 1% Triton X-100, 1 mM EDTA, complete protease  inhibitor cocktail). Samples were immunoprecipitated at 4 ºC overnight and washed six  times with ChIP wash buffer (20 mM Tris, pH 7.9, 500 mM NaCl, 2 mM EDTA, 1% Triton  X-100, 0.1% SDS) and once with TE buffer. Bound DNA was eluted in 1% SDS buffer  and reverse-crosslinked for 6 h at 65 ºC DNA samples were treated sequentially with   RNase A and Protease K and then purified with ChIP DNA Clean & Concentrator-5  (Zymo Research, D4014).The End-It DNA End-Repair Kit (Lucigen, ER81050) was used to convert purified DNA to 5′-phosphorylated, blunt-end DNA. After treated with Klenow Fragment (3'→5' exo-), barcodes (BIOO Scientific, 514120) were ligated with Quick Ligation kit (NEB, M2200). After size selection, 15 cycles of PCR were used to amplify the library.