RAPD-PCR and SSCP analysis for insect population genetic studies

Abstract

Population genetic studies require analysis of many individuals with multiple genetic markers. Prior to the development of the polymerase chain reaction (PCR) insect population geneticists relied almost exclusively on biochemical polymorphisms (Tabachnick and Black, 1996) as genetic markers. Random Amplified Polymorphic DNA (RAPD) and Single-Strand Conformation Polymorphism (SSCP) analysis are two PCR-based techniques developed in the last 5 years that allow a virtually unlimited number of genetic markers to be amplified in individual insects. Both techniques are simple, rapid, safe and relatively inexpensive. They offer the insect geneticist an unprecedented ability to examine many genetic markers simultaneously. This improves resolution of spatial and seasonal genetic differentiation among populations and increases the accuracy and precision of gene flow rate estimates. Monitoring segregation of markers distributed throughout a genome also allows for intensive examination of linkage disequilibrium. RAPD and SSCP markers have already been used in population genetic studies (Puterka et al., 1993; Tabachnick and Black, 1996; Apostol et al., 1996; Norris et al., 1996) and in genetic fingerprinting (Apostol et al., 1993,1994). They have also proven useful in detection and identification of cryptic species (Black et al., 1992; Wilkerson et al., 1993; Kambhampati et al., 1992; Ballinger-Crabtree et al., 1992; Hiss et al., 1994; Black and Munstermann, 1996).