Comparison of Control Region Sequencing and Fragment RFLP Analysis for Resolving Mitochondrial DNA Variation and Phylogenetic Relationships among Great Lakes Walleyes

Abstract

Direct sequencing of 513 base pairs from the control region and restriction fragment length polymorphisms (RFLP) in two fragments totaling 7.6 kilobases (fragment RFLP) that were amplified by polymerase chain reaction were used to assess mitochondrial DNA (mtDNA) variation in Great Lakes walleye Stizostedion vitreum. Our objective was to determine the effectiveness of these mtDNA markers in detecting genetic variation and resolving phylogenetic relationships among haplotypes previously identified by RFLP analysis of the entire molecule. The fragment RFLP analysis surveyed 554 base pairs and detected almost twice as many haplotypes as did the sequencing analysis. However, both approaches resulted in similar tree topologies and resolved the three major phylogenetic assemblages published in entire-molecule RFLP studies.