CUT&RUN in pantry moth testes, H4K16ac, replicate 3
Cells for CUT&RUN 51 were isolated from late 4th/early 5th instar Plodia larval testes by pulverization with a pestle in a 1.5 mL tube. ~500,000 cells were harvested per reaction. CUT&RUN was performed using the CUTANA ChIC/CUT&RUN Kit (EpiCypher) following the manufacturer's protocol. CUT&RUN libraries were generated using NEBNext Ultra II DNA Library Prep Kit for Illumina (New England Biolabs) with the following modifications in PCR amplification cycling parameters: initial denaturation at 98oC for 45 sec; 14 cycles of denaturation at 98oC for 15 sec and annealing/extension at 60oC for 10 sec; and final extension at 72oC for 1 min. Index primers used for CUT&RUN libraries generation were NEBNext Multiplex Oligos for Illumina (New England Biolabs). QC of CUT&RUN libraries was performed using Qubit and TapeStation. Libraries were sequenced on a Illumina NovaSeq 6000 with SP100 flow cell (50 bp paired-end).