GSM8797707: his-9/H3.1 reporter strain, rep 3, whole organism, fed, V5 ChIP; Caenorhabditis elegans; ChIP-Seq
Frozen worms were lysed by a pre-chilled biopulverizer (Biospec, #59013N). Pulverized samples were moved to a pre-chilled mortar and grinded vigorously with mortar and pestle (Fisher, #FB961B; #FB961L). Sample powder was moved to tubes and reconstituted in Nuclei Preparation Buffer (NPB) (10 mM HEPES pH 7.5, 10 mM KCl, 1.5 mM MgCl2, 0.1% Triton X-100) containing protease inhibitors (Promega, #G6521). Samples were cross-linked in 1.1% PFA (Fisher, # 50-980-487) for 4 minutes and quenched for 10 minutes with 125 mM (final concentration) Glycine (VWR, # 97061-128). Samples were washed in NPB + protease inhibitors and resuspended in lysis buffer (50 mM Tris pH 8.1, 10 mM EDTA, 0.25% SDS) plus protease inhibitors. A Diagenode Bioruptor Plus sonicator (#B01020001) and TPX tubes (Diagenode, #NC0065146) were used to sonicate samples to generate 400-500 bp DNA fragments. Input and IP sequencing libraries were generated using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (NEB, #E7645S) with barcodes unique to each of the samples (NEB, #E7335S; #E7500S).
GEO
GSM8797707_r1