3rd instar brains and imaginal disks, H3K27me3 input
3rd instar larvae
CNS and imaginal disks
Metadata from Sequence Read Archive
Fixed brains and imaginal discs were dissected from 10 third instar larvae. Sonicated chromatin incubated with anti ChIP was performed with 1:100 dilutions of anti-Pho (custom made) and anti-Ph antibodies (a kind gift from Donna Arndt-Jovin), 1:200 dilutions of anti-H3K27me3 (Millipore, 17-622) antibodies, and anti-Engrailed antibodies. Input ChIP DNA (30 ng) was blunt-ended and phosphorylated. A single 'A' nucleotide was added to the 3' ends of the fragments in preparation for ligation to an adapter that has a single-base 'T' overhang. The ligation products were purified and accurately size-selected by agarose gel electrophoresis. Size-selected DNA was purified and PCR-amplified to enrich for fragments that have adapters on both ends. The final purified product was then quantitated before cluster generation.