GSM4175898: Setdb1 young rep3; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
No description
Antigen
NA
Cell type
Cell type Class
No description
Cell type
NA
NA
NA
Attributes by original data submitter
Sample
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Snap-frozen mouse liver (100 mg) from wild type mice was used to prepare chromatin. Rabbit polyclonal antibodies specific to Setdb1 (Proteintech, 11231-1-AP), Kap1 (Abcam, ab10483), and anti-acetyl-histone-H3 specific to Lys-14 (Millipore, 07-353) were used for immunoprecipitation. Libraries were prepared according to Illumina's instructions. Briefly, DNAs were blunted with a combination of T4 DNA polymerase, Klenow polymerase, and T4 PNK, then a single 3′-end “A” base was added using Klenow exo (3′-to-5′ exo minus). multiplex adapters were then ligated to the ends of the modified DNA. After adapter ligation DNA was PCR amplified with Broad custom primers for 16 cycles and library fragments of ~280 bp (~150-bp mononucleosome insert plus ~130-bp adaptor and PCR primer sequences) were isolated using Pippin Prep agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on Illumina NextSeq 500 (Sedb1 & Kap1) and Illumina Hiseq 2500 (H3K14ac) following the manufacturer's protocols.