P-GSE45123-3 - P-GSE45123-3
For each biological replicate genomic DNA was generated from 15 parental or selected F2 flies using DNA Easy prep kit (Qiagen). To avoid RNA contamination, genomic DNA was treated with 1.5μl RNAse at 37°C for 30 min. Linear amplification of genomic DNA was performed using the REPLI-g midi kit to a concentration of ~ 1μg/μl. Five μg of amplified genomic DNA was fragmented with 1 U RQ1 DNase (Promega) for 2 min at room temperature (18–22°C) in standard buffer followed by incubation with 2ul EDTA at 65°C for 10 min, and digestion was confirmed on 3% agarose gels by the existence of sheared products with ~50–100 bp length.