GSM4191566: Input DNA for RNAPII; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Embryonic fibroblast
Cell type
MEF
Tissue
Embryonic Fibroblast
Lineage
primaryCells
Description
Mouse Embryonic Fibroblast
Attributes by original data submitter
Sample
source_name
primary mouse embryonic fibroblasts (MEFs)
strain
C57BL/6
developmental stage
embryo
genotype
wild-type
tissue
primary mouse embryonic fibroblasts (MEFs)
chip-antibody
none (input)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
After stimulating MEFs with TNF, MEF cells were washed with PBS and double cross-linked with 100mM disuccinimidyl glutarate/PBS solution (DSG, ThermoFisher Scientific) for 30-min followed by 1% methanol-free Formaldehye/PBS solution (ThermoFisher Scientific) for 15-min. Cross-linked cell pellets were quenched with 125mM Glycine for 5-min. Cell lysates were washed with PBS, incubated in lysis buffer, and sonicated to obtain DNA fragments of 250 to 1000 bp. Cells were sonicated with Diagenode Bioruptor 300 sonication system. DNA fragments were incubated with 5 ug anti-Pol II antibody (Santa Cruz Biotechnology, sc-899) overnight and immunoprecipitated DNA fragments were subjected to reverse cross-linking and cleaned up by AMPure XP magnetic beads. 5ng of input or ChIP DNA were subjected for library synthesis without size selection using NEBNext Ultra DNA Library Prep Kit for Illumina (New England BioLabs) according to manufacturer's instruction. ChIP DNA libraries were quantified using Qubit 2.0 Fluorometer.