GSM1930760: MCF-10A wildtype BRG1 ChIPseq Replicate1 Pulldown; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Unclassified
Antigen
Unclassified
Cell type
Cell type Class
Breast
Cell type
MCF 10A
Primary Tissue
Breast
Tissue Diagnosis
Fibrocystic Disease
Attributes by original data submitter
Sample
source_name
Mammary epithelial cell line
cell line
MCF-10A
cell type
Mammary epithelial cell line
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
~1x107 parental MCF-10A cells were crosslinked with formaldehyde at room temperature for 10 minutes. Then, the cells were lysed using lysis buffer A (50mM HEPES, 140mM NaCl, 1mM EDTA pH=8, 10% Glycerol, 0.5% NP-40, 0.25% Triton X-100), and the residual cytoplasmic protein was removed using lysis buffer B (10mM Tris-HCl pH=8, 200mM NaCl, 1mM EDTA, 1mM EGTA). The nuclear fraction was released using lysis buffer C (10mM Tris-Hcl pH=8, 100mM NaCl, 1mM EDTA, 1mM EGTA, 0.1% Sodium Deoxycholate, 0.5% N-lauroylsarcosine). The chromatin was then sheared by using a Bioruptor instrument in high setting, 30' on , 30' off for 5 minutes for 5 cycles. The pull-down was performed by using the BRG1 antibody (Santa Cruz #G-7). Samples were washed three times with RIPA buffer (Tris-HCl pH=8, 150mM NaCl, 1mM EDTA, 1% NP-40, 0.25% Sodium deoxycholate, 0.1% SDS) and were eluted. The pull-down and input control sequencing libraries were generated by using the NEXTflex Rapid DNA Sequencing Kit (Bioo Scientific #5144-02) and were sequenced by using SE100 reads with a HiSeq 2000 instrument.