GSM3058480: HaCaT DMSO Input DNA; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Epidermis
Cell type
HaCaT
Primary Tissue
Skin
Tissue Diagnosis
Normal
Attributes by original data submitter
Sample
source_name
HaCaT
cell line
HaCaT
cell type
Human immortalized keratinocytes
chip antibody
none
culture media
+DMSO
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Cells grown on 10cm plates were crosslinked with 1% formaldyhyde in culture media. Cross-linked chromatin was collected and flash frozen before sonication with the Ultrasonic Cell Crusher. Lysates were clarified and protein-DNA complexes were immunoprecipitated with H3K27me3 antibody. Libraries were prepared according to standard Illumina protocols. Libraries were prepared according to NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB #E7645). Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E.coli DNA Pol I Large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenowfragment (32 to 52 exominus) and dATP to yield a protruding 3-'A' base forligation of Illumina's adapters which have a single 'T' base overhang at the 3' end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of 200-500bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was capturedon an Illumina flowcell for cluster generation. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols.