GSM1545045: Control H3K4me3; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Histone
Antigen
H3K4me3
Cell type
Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA
Attributes by original data submitter
Sample
source_name
WW6 cells
cell type
mouse embryonic stem cells
strain
~20% C57/B6J, ~75% 129/Sv and ~5% SJL
treatment
transfected cells with mock control construct
chip antibody
H3K4me3 (Abcam ab8580, lot GR95252-1)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
3 million cells crosslinked in 1% formaldehyde were lysed with Farnham and RIPA cold lysis buffers. Following collection, the crude nuclear preparation was processed in a Bioruptor at the high setting for a total of 30 minutes, in cycles of 30 seconds on/30 seconds off. The chromatin was collected by centrifugation and incubated with 5 µg of H3K4me3 antibody overnight. The immunoprecipitated samples were end-filled using a combination of T4 DNA polymerase and T4 polynucleotide kinase , 3’ -terminal-A extended with Klenow 5'-3' exo minus, and ligated to pre-annealed TruSeq indexed Illumina adapters. Libraries were amplified using Illumina primers and gel extracted for size selection and primer-dimer removal. Before sequencing, libraries were tested using the BioAnalyzer to test quality in terms of size and primer-dimer depletion.