GSM8457987: [ChIP-Seq] biotin_CHD8_12h_rep2; Mus musculus; ChIP-Seq
Crosslinked cells were lysed in ChIP SDS lysis buffer containing 1x protease inhibitor cocktail and PMSF, then sonicated to achieve a chromatin sized of 200–400 bp. Supernatant were diluted 9-fold with dilution buffer and then precleared with Dynabeads protein A/G (1:1 mixed) for 1 hour at 4 °C and immunoprecipitated using 4 μg antibodies for 12 hours and 50 μl Dynabeads protein A/G (1:1 mixed) for 3 hours. Immune complexes were washed with the following buffers: low salt wash buffer, high salt wash buffer , LiCl wash buffer and TE buffer. ChIPed DNA was reverse-crosslinked. ChIP-seq libraries were constructed using the VAHTS Universal DNA Library Prep Kit for Illumina®V3 according to the manufacturer's instructions.