GSM1842756: mES Nanog 24h 2i r2; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Unclassified
Antigen
Unclassified
Cell type
Cell type Class
Pluripotent stem cell
Cell type
ES cells
NA
NA
Attributes by original data submitter
Sample
source_name
KH2 mouse embryonic stem cells
cell type
mouse embryonic stem cells
culture condition
2i for 24 hours_v1
antibody epiptope
Nanog
antibody-vendor
Cosmo Bio Ltd
antibody-catalog-nr
REC-RCAB0002P-F
antibody-lot
20100713
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
RNA-Seq: Polyadenylated RNA was isolated using Olig dT beads (Invitrogen). RNA was fragmented to a size of 200-600 base pairs, followed by ligation to RNA adapters using T4 RNA Ligase (NEB). Adapter primers were used for cDNA synthesis, followed by RNA degradation. Each cDNA library was ligated to a DNA adapter, used for barcoding and PCR enrichment of the library. Libraries were pooled and sequenced on the Illumina HiSeq 2000. ChIP-Seq: Cells were crosslinked in 1% formaldehyde for 15 minutes at room temperature, with constant agitation, followed by quenching with 125mM Glycine for 5 minutes at room temperature. Nuclei were isolated and chromatin was sheared using Branson sonifier until the majority of DNA was in the range of 200-700 base pairs. Chromatin was incubated with antibody overnight at 4°C, with constant agitation. Immunoprecipitation of antibody-protein complexes was completed using Protein A or Protein G Dynabeads (Invitrogen) for 2-3 hour at 4°C, with constant agitation Immunoprecipitated DNA was end repaired using the End-It DNA End-Repair Kit (Epicentre), extended using a Klenow fragment (3’-5’ exo) (NEB), and ligated to sequencing adapter oligos (Illumina). Each library was then PCR-amplified using PFU Ultra II Hotstart Master Mix (Agilent), and a size range of 300-600 was selected for sequencing on the Illumina HiSeq 2000.