progeny of mat-a-tubGal4/controlRNAi mothers crossed to their siblings (Bloomington stocks: 7062 & 36304)
tissue
blastoderm embryos c14
technique
Mnase-ChIP-seq
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
ChIPseq: Dechorionated embryos were fixed using 0.1% Formaldehyde for 15min at RT. After fixation, they were hand picked for the desired developmental stage and frozen at -80oc until further use. Nuclei were isolated using short covaris sonication followed by chromatin digestion using Micrococcal nuclease and 10 cycles of sonication in a Bioruptor Pico. The lysate was clarified by centrifugation before immunoprecipitation, washes, elution by reverse crosslinking at 65deg for 16h. After RNaseA and proteinaseK treatment, the DNA from input and immunoprecipitation was purified using phenol-chloroform extraction and ethanol precipitation. NEB Next Ultra II