Experimental selection of hypoxia-adapted Drosophila melanogaster
Genomic DNA was isolated from a pool of 100 male and 100 female adult flies collected from hypoxia-selected populations or generation matched control populations by standard phenol:chloroform extraction followed by treatment with DNase-free RNase. DNA quality was assessed by spectrophotometry (260/280 and 260/230) and gel electrophoresis. A total of 3µg was sheared DNA (Covaris, Woburn, MA) and used to construct library for paired-end sequencing. The DNA fragments were subject to end repair using End-IT DNA End-Repair Kit (Epicentre, Madison, WI) and then ligated to Illumina PE adapters. The adapter-ligated products were purified on Qiaquick spin columns (Qiagen Inc., Valencia, CA) and PCR-amplified with high fidelity DNA Polymerase in 12 cycles using Illumina’s PE primer set (Illumina, San Diego, CA). Cluster generation was performed using the Illumina cluster station and cluster generation kit v2.