title: GSM1148116: ASC_y_H3K36me3_ChIPSeq; Mus musculus; ChIP-Seq
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.