GSM3670062: Cal27 H3K27ac ChIP seq; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Histone
Antigen
H3K27ac
Cell type
Cell type Class
Others
Cell type
CAL 27
Primary Tissue
Tongue
Tissue Diagnosis
Carcinoma Squamous Cell
Attributes by original data submitter
Sample
source_name
Tongue
cell line
Cal27
tissue
Tongue
tumor type
HNSCC
chip antibody
anti-H3K27Ac (Active Motif, #39133)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Chromatin was sheared in diluted lysis buffer (1% SDS, 50mM Tris-HCl, 20mM EDTA, 1x protease inhibitor) to 200-500bp using a Covaris M220 Focused-Ultrasonicator with the following parameters: 10 minutes, peak incident power 75, duty factor 10%, 200 cycles/burst. 3ug antibody anti-H3K27Ac (Active Motif, #39133) was incubated with Dynabeads Protein G beads (Invitrogen, #10004D) for 4 hours. Beads were washed 3 times with 5mg/ml BSA, and incubated with 50% crosslinked chromatin (500ul) and 50% ChIP master mix (1% Triton-X, 0.1% DOC, 1X protease inhibitor, 1x TE) at 4°C overnight. 10% of the chromatin was not exposed to antibody and was used as control (input). Immunoprecipitated DNA was washed with RIPA buffer (50mM Hepes, 1% NP-40, 0.7% DOC, 0.5M LiCl, 1mM EDTA, 1x protease inhibitor) and eluted from the beads in elution buffer (10mM Tris, 1mM EDTA, 1% SDS). Along with input, immunoprecipitated DNA was reverse crosslinked at 65°C overnight, and purified via Phenol:Chloroform extraction DNA libraries were prepared with either the TruSeq ChiP Library Prep Kit (Illumina, #IP-202-1012) or the Accel-NGS 2S Plus DNA Library kit (Swift Bioscience, #21024) and sequenced using 75 bp single-end sequencing method on an Illumina Hi-seq 4000