We performed CUT&RUN in wing disc tissue, as described in (109 (link)), modified from (49 (link)). For each replicate, 20 WL3 imaginal wing discs were used. Two wing discs of Drosophila yakuba were also included with each replicate but were not used in downstream analyses. For the H3K27me3 experiment, αH3K27 (Cell Signaling Technology, no. 9733, 1:100) and protein AG-MNase (1:100; UNC core) were used. For the H3.3 experiment, αH3.3 (H3F33B, 1:100; Abnova, no. H00003021-M01) and protein A-MNase (gift of S. Henikoff) were used.
The ThruPLEX DNA sequencing kit was used for the library preparation. The manufacturer’s protocol was followed until the amplification step. After the addition of indexes, 16 to 21 cycles of 98˚C, 20 s and 67˚C, 10 s were run. DNA library purification was done using AMPure XP beads. Libraries were sequenced on an Illumina NextSeqP2.
The ThruPLEX DNA sequencing kit was used for the library preparation. The manufacturer’s protocol was followed until the amplification step. After the addition of indexes, 16 to 21 cycles of 98˚C, 20 s and 67˚C, 10 s were run. DNA library purification was done using AMPure XP beads. Libraries were sequenced on an Illumina NextSeqP2.
