PT - JOURNAL ARTICLE AU - John K. Kelly AU - Kimberly A. Hughes TI - An examination of the evolve-and-resequence method using <em>Drosophila simulans</em> AID - 10.1101/337188 DP - 2018 Jan 01 TA - bioRxiv PG - 337188 4099 - http://biorxiv.org/content/early/2018/10/12/337188.short 4100 - http://biorxiv.org/content/early/2018/10/12/337188.full AB - We develop a set of analytical and simulation tools for Evolve-and-Resequence (E&amp;R) experiments and apply them to a new study of rapid evolution in Drosophila simulans. Likelihood based test statistics applied to pooled population sequencing data suggest parallel evolution of 138 polymorphisms (SNPs) across the genome. This number is reduced by orders of magnitude from previous studies (thousands or tens of thousands), owing to differences in both experimental design and statistical analysis. Whole genome simulations calibrated from several Drosophila genetic datasets support the contention that the observed genome-wide response could be generated by as few as 30 loci under strong directional selection, with a corresponding hitch-hiking effect. Finally, the SNPs that showed strong parallel evolution in the novel laboratory environment exhibit an (initial) allele frequency spectrum indicative of balancing in nature. These loci also exhibit elevated differentiation among natural populations of D. simulans.