GSM2648437: hp Infected WT 7480 Bcl11b ChIP; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
Bcl11b
Cell type
Cell type Class
Blood
Cell type
CD4+ T cells
NA
NA
Attributes by original data submitter
Sample
source_name
CD4+ T-cells
strain
C57BL/6
chip antibody
anti-Ctip2 (Bcl11b) (abcam ab18465)
genotype
Wild type
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Isolated cells were fixed with 1% formaldehyde in PBS for 10 minutes, then quenched with glycine. After washing, DNA was extracted using the Illumina SimpleChIP Enzymatic ChIP kit (#9003S). Briefly, nuclei were isolated by incubating in 1ml buffer A for 10 min on Ice, followed by centrifugation and resuspension in 1 ml Buffer B. Chromatin was digested for 20 min at 37oC in 1 ml buffer B with 2 ul 1:10 diluted micrococcal nuclease. Digestion was stopped with EDTA. Nuclei were lysed by sonication, centrifuged to clear lysates, and the supernatant containing the DNA was transferred to a fresh tube for overnight antibody complexing. Next day, protein A dynalbeads were added to each tube, incubated, washed, and isolated with magnet. DNA complexes were then released from the beads, decrosslinked, and phenol:choloroform:isolamyl alcohol extracted for library prep. DNA libraries were prepared using NEBNext Ultra II DNA Library Prep Kit for Illumina. Briefly, DNA ends were repaired, followed by ligation of adapter sequences. DNA was cleaned with AMPure XP beads, and then DNA was PCR amplified with 7-8 cycles. After final cleaup with AMPure XP Beads, libraries were quantified using Agilent Bioanalyzer high sensitivity DNA kit and were subsequently sequenced.