GSM7140425: C. elegans, HIF-1a, IP, Bio rep1; Caenorhabditis elegans; ChIP-Seq
Worms were harvested and cross-linked in 2% formaldehyde at 21°C for 30 minutes. Nuclear lysates were sonicated using a Branson sonifer microtip to fragment the chromatin to 200-800 bp. Immunoprecipitated protein-DNA complexes were captured on protein A-Sepharose beads (Sigma) and eluted in elution buffer (1% SDS and 100 mM NaHCO3) at 65°C for 30 minutes. Following RNase treatment and cross-link reversal, the ChIP DNA was purified with the Qiagen MinElute Kit for sequencing in parallel with the corresponding input DNA. The ChIP-seq library preparation and sequencing were performed by the Iowa State University DNA facility. In brief, NEXTflex™ ChIP-Seq Barcodes kit (Illumina compatible) (BIOO Scientific Corp., cat. no. 514123) was used to prepare multiplexed single-end genomic DNA libraries. The gel slices corresponding to the 200-300 bp maker were cut and purified. The purified DNA was amplified and sequenced in a single flow cell on the IlluminaHiSeq 2000 platform. The length of single-end reads was 50 bp.