pancreatic adenocarcinoma cancer Tet-on KrasG12D cell line
genotype/variation
tet-on_KrasG12D;KO_Cdkn2a;KO_Smad4;KO_Trp53
chip antibody
anti-Foxa2 (Cell Signaling, #8186)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
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)