GSM1148117: ASC y input DNA; Mus musculus; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
Input control
Antigen
Input control
Cell type
Cell type Class
Muscle
Cell type
Skeletal muscle
NA
NA
Attributes by original data submitter
Sample
source_name
hindlimb muscles
strain
C57BL/6
gender
male
age
2-3 months
sorting markers
VCAM+ / CD31- / CD45- / Sca1-
chip antibody
none
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
FACS-sorted cells were crosslinked with 1% formaldehyde at room temperature with gentle agitation for 10 minutes. The crosslink was terminated by Glycine at a final concentration of 0.125 M. Cells were then washed three times with ice-cold PBS and stored at -80°C until a sufficient number of cells had been obtained. ChIP was performed following standard ChIP protocols with the following major modifications: 10E6 cells were resuspended in 200 ml lysis buffer and sonicated to obtain DNA fragments from 150 to 500 bp in size. Each ChIP reaction was performed with 10E6 cells and 5 ug of antibody. The ChIP DNA was size-selected for fragments between 150-400 bp by 2% low-melt agarose gel electrophoresis (NuSieve). A library for deep sequencing was generated with Illumina ChIP-seq Sample Prep Kit (Part# IP-102-1001) with the following modification to the standard protocol: The adaptor solution was diluted 50 times for ligation. 15 cycles of PCR amplification were performed immediately following adaptor ligation. Size-selection of fragments between 200-400 bp was performed after PCR amplification. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer IIx following the manufacturer's protocols.