GSM5021152: RWPE1-Vector ERK2 ChIP; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
MAPK1
Cell type
Cell type Class
Prostate
Cell type
RWPE-1
Primary Tissue
Prostate
Tissue Diagnosis
Normal
Attributes by original data submitter
Sample
source_name
RWPE1
cell type
normal prostate epithelial cell line
cell line
RWPE1
retrovirus
Retrovirally transduced and selected to express pQCXIH
chip ab
ERK2 (D-2 SCBT)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Nuclear lysates were incubated with 5uL of indicated antibody and 50uL of Dynabeads for 4 hours in dilution buffer(20mM Tris, 2mM EDTA,150mM NaCl, 1% Triton X-100, 2 mg/mL BSA) followed by 4 washes with wash buffer(20mM Tris, .25% NP-40, .05% SDS, 2mM EDTA, 250mM NaCl). ChIPs were then incubated with RNAse A (5Prime) and Proteinase K (Sigma) followed by DNA extraction with Qiagen PCR purification kit. Total Cellular RNA was extracted using Qiagen RNeasy kit followed DNAse treatment (Turbo) and poly-A selection using oligo dT beadsd (Ambion). Poly-A selected RNA was then reverse transcribed using random hexamer primers and SuperScript III. Double stranded DNA was then produced from cDNA fragments using DNA ligase and DNA polymerase. Isolated DNA was sonicated for 30 minutes (30sec on / 30 sec off) followed by End Repair with T4 DNA polymerase, Klenow DNA polymerase, and T4 PNK. End repaired DNA was 3' Adenylated with Klenow exo (3' to 5') followed by ligation of Illumina TruSeq adaptors with T4 DNA ligase. Ligated DNA fragments were subject to 15 cycles of PCR with Q5 polymerase followed by size selection with AMPure XP beads.