GSM9616231: fed C. elegans L1, LIN-15B ChIP-seq, lin-35, rep2; Caenorhabditis elegans; ChIP-Seq
Worm pellets were flash-frozen in liquid nitrogen and ground to powder. Crosslinking was performed using 1.5 mM EGS followed by 1% formaldehyde. Chromatin was extracted and sonicated to an average fragment size of ~250 bp. Immunoprecipitation was performed using antibodies against LIN-15A (Q2015), LIN-15B (Q2330), and LIN-35 (Q2001). ChIP DNA was converted into sequencing libraries using a modified Illumina TruSeq protocol. DNA fragments were end-repaired and purified using AMPure XP beads. Fragments were A-tailed using Klenow (3′→5′ exo−), followed by ligation of Illumina TruSeq adapters. Adapter-ligated DNA was purified and amplified by PCR using KAPA HiFi HotStart polymerase with Illumina universal primers. Libraries were size-selected using AMPure XP beads to enrich for appropriate fragment sizes and remove adapter dimers. Final libraries were quantified using Qubit and assessed for quality and fragment size distribution using an Agilent TapeStation.