GSM1826440: Input DNA rep1; 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
HMEC
Primary Tissue
Breast
Tissue Diagnosis
Normal
Attributes by original data submitter
Sample
source_name
Human mammary epithelial cells (HMEC)
treatment
untreated
shRNA
none
chip antibody
none
cell type
HMEC
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
For ChIP-seq: Cells were crosslinked with 2 mM disuccinimidyl glutarate (DSG) for 45 min and then 1% formaldehyde for 15 min at room temperature. Following cell lysis, chromatin was sheared using a Misonix 3000 probe sonicator (30 x 30 sec, 30 sec off). p65-bound chromatin was then immunoprecipitated using 8 micrograms anti-p65 (sc-372/C-20X) antibody. Following washing and reverse crosslinking, chromatin was purified using phenol-chloroform extraction. For RNA-seq: total mRNA was isolated using RNeasy Kit (Qiagen). DNA and mRNA libraries were created following standard Illumina protocols with PolyA capture for mRNA. ChIP-seq and RNA-seq; 2-lane flow cell (single-end 50 bp sequencing)