title: Low Feeder Population 2, generation 10
design: Starting from an AIP base population derived from the DGRP, we scored 100 virgin females and males, and selected the 20 individuals of each sex with the highest food consumption to establish high feeding selection lines. In each subsequent generation, we selected the 20 highest males and females from 100 scored of each sex as parents of the next generation.   We then prepared a DNA library by pooling at least 100 female flies from mix of base populations at generation 9 or 10 of mass selection for low feeding behavior. Flies were flash frozen on clean CO2 pads and homogenized using TIssuelyser (Qiagen, Inc.) in lysis buffer. Genomic DNA was extracted using magnetic beads and fragmented to 300-400bp using Covaris S220. Fragmented DNA was used to produce barcoded DNA libraries using NEXTflex™ DNA Barcodes (Bioo Scientific, Inc.) with an Illumina TrueSeq compatible protocol. Libraries were quantified using Qubit dsDNA HS Kits (Life Technologies, Inc.) and Bioanalyzer (Agilent Technologies, Inc.) to calculate molarity. Libraries were then diluted to equal molarity and re-quantified, and all samples were pooled together. Pooled library samples were quantified again to calculate final molarity and then denatured and diluted to 14pM. Pooled library samples were clustered on an Illumina cBot and sequenced on two Hiseq2500 high throughput lanes using 125 bp paired-end v4 chemistry.