title: Japan_D.melanogaster
design: DNA was extracted from single flies using DNeasy kits (Qiagen) according to the manufacturer's protocol. Each DNA extraction was amplified using GenomiPhi (GE Healthcare, and the four DNA extractions were quantified using SYBR green fluorescence, and then combined in an equimolar mixture. PCR (Qiagen, LongRange PCR) used to amply the regions of interest (5Kbp amplicons). PCR products from each population pool of 4 chromosomes were quantified using the brightness of bands on an agarose gel and mixed in approximately equimolar amounts. To remove unincorporated nucleotides, primers and PCR product of the incorrect length, this mixture was then run on an agarose gel, the band excised, and the DNA purified from the gel slice.