GSM6267710: Worker H3K27me3 replicate 2; Apis mellifera; ChIP-Seq
Extracts from sonicated nuclei were cleared by centrifugation (20000×g; 10 min), histone-DNA complexes were concentrated using an Amicon centrifugal filter (Millipore) and split into 50 μl aliquots. Each aliquot was diluted ten-fold with ChIP dilution buffer (0.01% SDS; 1.1% Triton X-100; 1.2 mM EDTA; 16.7 mM Tris-HCl pH 8.0; 167 mM NaCl; protease inhibitors cocktail) and histone-DNA complexes were immunoprecipitated with antibodies overnight. Libraries were prepared with the NEBNext Ultra II Library Prep Kit for Illumina (NEB, E7645S) according to the manufacturer's protocol. Insert sizes were selected for 250 bp with NEBNext Sample Purification Beads (NEB, E7104S). Libraries were PCR amplified using NEBNext Multiplex Oligos for Illumina (Index Primers Set 1) (NEB, E7335S). The number of PCR cycles was selected based on the amount of input DNA. Library quality was verified by running an Agilent High Sensitivity DNA chip (Agilent, 5067-4626) on a 2100 Bioanalyzer (Agilent Genomics).