GSM2643616: ChIPseq.HumanPRDM9 HA.Input.ProtocolC; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Kidney
Cell type
293
Primary Tissue
Kidney
Tissue Diagnosis
Normal
Attributes by original data submitter
Sample
source_name
HEK293T cells
cell line
HEK293T (ATCC CRL-3216)
transfected with
HumanPRDM9(B)_HA
chip antibody
None
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP was performed as described in Gupta-Hinch et al. 2014, with modifications. Crosslinking was performed in 1% formaldehyde for 5 minutes. Input chromatin was "pre-cleared" to remove chromatin bound non-specifically by the beads. For each sample, 50 ul of equilibrated magnetic beads were resuspended in 100 ul PBS/BSA and added to the chromatin samples for pre-clearing for two hours at 4C with rotation. Beads were removed, and 100 ul of pre-cleared chromatin was set aside for the input control. 5 ul ChIP-grade rabbit polyclonal antibody was added to the remaining pre-cleared chromatin and incubated overnight at 4C with rotation. 50 ul beads were washed and resuspended as before, then incubated with the chromatin samples for two hours at 4C with rotation. After washing and decrosslinking, samples were further incubated with 80 ug RNAse A at 37C for 60 minutes and then with 80 ug Proteinase K at 55C for 90 minutes. Libraries were prepared for sequencing using standard Illumina protocols by the Oxford Genomics Centre.