Studies on Genotyping of Drosophila White-one Mutation using the Polymerase Chain Reaction: Use of Frozen Flies, Non-toxic Stains, and Direct Polymerase Chain React

Abstract

To optimize the laboratory protocol for the PCR-based assay genotyping of Drosophila melanogaster white-one mutation for an undergraduate biology course, this research project investigates how the freezing of Drosophila adults affects the quality of PCR products, and explores direct PCR and non-toxic DNA stains as alternatives to standard PCR and toxic ethidium bromide, respectively. Objectives of this project are to study effects of length of freezing time and multiple freeze/thaw cycles on the quality of PCR products and compare alternative methods to current methods in product quality. This research project has implications in terms of storage and handling of Drosophila adults in class and serves as a starting point in the pursuit of more consistent, quicker, and safer methods in the future.