GSM2698024: WT Dnmt3a1 Input DNA r2; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA
Attributes by original data submitter
Sample
source_name
embryonic stem cells
strain
129S4/SvJae
genotype
wild type
chip antibody
none
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
5×106 cells were fixed for 10 minutes with 1% formaldehyde, followed by chromatin extraction and sonication to generate 200-500bp fragments. Ten percent of chromatin is kept as Input. Immunoprecipitation was performed overnight at 4°C using streptavidin-M280 (Thermo Fisher, 11205D) magnetic beads. ChIP washes were performed in the following order: 2% SDS, 1×high-salt buffer, 1×LiCl buffer, 1×TE buffer, 1×TE buffer 0.2% Triton X-100. Cross-linking was reversed with the addition of elution buffer (20 mM Tris-HCl, pH7.5, 5 mM EDTA, 50 mM NaCl, 1% SDS, 50 μg/mL Proteinase K) to washed beads and incubation at 68°C with rotation for three hours. DNA was purified with MinElute PCR purification kit (Qiagen, 28004). For ChIP-Seq, sequencing libraries were prepared using ThruPLEX DNA-seq 48D kit (Rubicon Genomics, R400406) following standard protocols.