GSM1411216: MCF7 Input single end ChIP-seq, rep2; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Breast
Cell type
MCF-7
Primary Tissue
Breast
Site of Extraction
Pleura
Tissue Diagnosis
Adenocarcinoma
Attributes by original data submitter
Sample
source_name
MCF-7, untreated, ChIP input
cell line
MCF-7
cell type
mammary adenocarcinoma cell line
treatment
none
capture oligo
N/A
antibody
none
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
For ChIP-seq, 10^8 MCF-7 cells (ATCC HTB-22) were crosslinked for 10 minutes with 1% formaldehyde. After nuclei isolation and sonication, sheared chromatin was incubated overnight with protein A beads bound to PSF antisera (clone 39-1; Santa Cruz Biotechnology, sc-101137, Lot # L1012). After incubation, beads were extensively washed and bound DNA was eluted through incubation with proteinase K at 55 degrees C for 1 hour. Eluted DNA was subjected to crosslink reversal overnight at 65 degrees C. DNA was purified using ethanol precipitation and subjected to library construction. Genomic DNA isolated by ChIP enrichment, as well as input DNA, was fragmented using sonication. Sonicated DNA was then subjected to end-repair, A-tailing, ligation to universal adapters, amplification with indexed primers, and size selection using SPRI beads.