H3K27me3 ChIP-seq in hTERT-RPE1 treated with HBiX1 siRNA
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Histone
Antigen
H3K27me3
Cell type
Cell type Class
Neural
Cell type
hTERT RPE-1
Primary Tissue
Eye
Tissue Diagnosis
Normal
Attributes by original data submitter
Sample
sample_name
DRS003046
sample comment
H3K27me3 ChIP-seq in hTERT-RPE1 treated with HBiX1 siRNA (mouse monoclonal; CMA323)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIP-seq libraries were prepared according to the manufacturer's instructions (Illumina). Briefly, immunoprecipitated DNA was end-repaired using a mix of Klenow DNA polymerase, T4 DNA polymerase and T4 polynucleotide kinase (NEB), tailed with an 'A' base using Klenow Fragment 3'-5' exo minus (NEB), and ligated with the Illumina single-end adaptor, using a DNA ligation Kit (TaKaRa). Adaptor-ligated fragments of approximately 300 bp were purified using the E-gel SizeSelect system (Life Technologies), and were subjected to 18 cycles of PCR amplification using KOD FX polymerase (TOYOBO). To remove the remaining PCR primers, the amplified products were further purified using AMPure XP Kits (Beckman Coulter). Libraries were quantified using a BioAnalyzer 2100 with a High Sensitivity DNA Kit (Agilent).