Application of the IRAP method for analysis of genetic diversity in perch (Perca fluviatilis) populations

Abstract

Studies of allele frequencies and genetic diversity are important to understand evolutionary processes of populations. Different types of retrotransposons-based molecular markers are universal and informative, therefore they are vide used for genetic analysis in animals, yeasts and plants. The goal of the study was to determine applicability of IRAP (Inter-Retrotransposon Amplified Polymorphism) marker for the studies of the genetic diversity of perch (Perca fluviatilis) populations from Latvian and Lithuanian lakes. On the first stage DNA were extracted from four perch blood and tissue samples and used for following PCR reaction with 26 primers, which were previously successfully used in the analysis of different animal and plant species. PCR products were tested in 1.7% agarose gel for 4.5 hours (80 V). On the second stage primers 2080, 2081 and 2239 with high level of polymorphisms were applied for further analysis, included 132 DNA samples from five Latvian lakes and 47 DNA samples from four Lithuanian lakes. PCR products were separated by the longer run in 1.7% agarose gel (15 hours, 50 V). Three used primers showed good applicability for analysis of perch genetic polymorphisms: 50 loci were revealed by the primer 2080, 42 loci – by primer 2081, and 35 loci – by primer 2239. The number of polymorphic loci in five Latvian and four Lithuanian populations was 119, and the percentage of polymorphic loci was 93.7%.