GSM8634630: TRF2, IP; Homo sapiens; ChIP-Seq
HCT116 (7 × 10^5) cells were plated and, after 72h, fixed with 1% of formaldehyde. Nuclei were lysed with SDS lysis buffer and lysates were sonicated by using the biorupter (Diagenode Inc., NXT-Dx Belgium). The average length of sonicated chromatin was around 300–400 bp. For each IP, 100 μg of chromatin and 5 μg of anti-TRF2 (NB110-57130, Novus) antibodies were used. ChIP samples were quantified by the Agilent 2100 Bioanalyzer System (Agilent Technologies) followed by library preparation for sequencing. DNA libraries were generated using the MicroPlex Library Preparation Kit protocol on the IP-Star Compact (Diagenode) apparatus following the manufacturer's instructions. Libraries were quantified using the KAPA Library Quantification Kit (KapaBiosystems) followed by sequencing of 1.8 pM per library in a NextSeq500 illumina system.