ChIP-seq of S. cerevisiae derivative yGWG605: NG dCas9-2xN with Targeting sgRNA, 0nM Beta-Estradiol, IP
Chromatin from formalin-crosslinked yeast was randomly sheared by sonication to generate predominantly 200-300bp DNA fragment lengths and treated with anti-HA antibody to capture fragments bound by HA-tagged (d)Cas9(-2xN). DNA was column-purified (MinElute, Qiagen) after elution from antibody-conjugated beads and de-crosslinking. Sequencing libraries were prepared from purified DNA samples using the ThruPLEX DNA-seq kit to introduce Illumina adapters and dual-indexing for multiplexed, paired-end (51 x 2) sequencing that read the ends of each DNA fragment.