title: GSM8646276: GFP::DMD-3, L4 males (strain DF301), Replicate 2, (Input); Caenorhabditis elegans; ChIP-Seq
library_construction_protocol: Samples were cross-linked by adding formaldehyde (2%) and incubating on a rocker at room temperature for 30 minutes. Glycine was added to a final concentration of 125mM and incubated at room temperature for 5 minutes.samples were dounce homogenized in FA-buffer (50 mM HEPES/KOH pH 7.5, 1 mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate; 150 mM NaCl). Sarkosyl (0.1% sodium lauroyl sarcosinate) was added to samples before sonication. Samples were sonicated to enrich chromatin fragments between 200 and 800 bp in length ChIP-DNA samples were ligated to barcode adaptors from the NEXTflex ChIPseq Barcodes kit (Bioo Scientific) using the NEXTflex ChIP-seq kit (Bioo Scientific) following the manufacturer's protocol with the following alterations: the initial size selection step (step B1) was skipped, the number of PCR cycles in the amplification step was 15. After amplification, 200-600 bp fragments were size-selected using SPRIselect beads (Beckman Coulter).