GSM1477662: Input DNA DCR Flox input-2; 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
mouse embryonic stem cells
cell type
mouse embryonic stem cells
strain
C57BL/6
genotype
DCR Flox
chip antibody
none
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
For ChIP 20 million ES cells were fixed, lysed and sonicated to 200 base pairs. Then, histone-DNA complexes were isolated with antibody. For RNA-Seq RNA was isolated from ~10^6 cells using the RNAzolRT reagent Libraries were prepared according to Illumina's instructions accompanying the DNA Sample Kit. Briefly, DNA was end-repaired using a combination of T4 DNA polymerase, E. coli DNA Pol I large fragment (Klenow polymerase) and T4 polynucleotide kinase. The blunt, phosphorylated ends were treated with Klenow fragment (32 to 52 exo minus) and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~200-400 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols. For RNAseq libraries were prepared either with the TruSeq RNA sample prep kit or the Directional RNA Sample prep kit (Illumina) and sequenced on a GAIIx.