GSM3416690: DCM5 Input; Homo sapiens; ChIP-Seq
Cardiomyocyte nuclei were isolated by FACS using anti PCM1 antibody. The nuclei were crosslinked using 1% formaldehyde for 2 min at room temperature. Crosslinked nuclei were fragmented using Bioruptor Pico (Diagenode) to achieve frgament size of 200-500bp. A portion of nuclear extract was kept aside to serve as input. Immunoprecipitation was carried out using  anti LMNA A/C antibody. Immunecomplexes were washed, eluted the DNA and reverse crosslinked at 65 O/N. the DNA was precipitated after phenol chloroform extraction by glycogen. ChIP libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit for Illumina (New England BioLabs, Inc.) following the manufacturer's protocol. Sequencing was done  on an Illumina HiSeq3000 for sequencing using a Single Read 50 bp run.