Capillary electrophoresis of an 11-plex mtdna coding region SNP single base extension assay for discrimination of the most common caucasian HV1/HV2 mitotype

Abstract

The typing of single nucleotide polymorphisms (SNPs) located throughout the human mitochondrial genome assists in resolving individuals with an identical HV1/HV2 haplotype. A set of 11 sites which were selected for distinguishing individuals of a common Western European Caucasian HV1/HV2 mitotype was incorporated into a single base extension (SBE) assay. The assay was optimized for multiplex detection of sequence polymorphisms at positions 3010, 4793, 10211, 5004, 7028, 7202, 16519, 12858, 4580, 477, and 14470 in the mitochondrial genome. PCR primers were designed to allow for multiplex amplification of unique regions in the mitochondrial genome followed by an 11-plex SBE reaction using the SNaPshot ® reagent kit. Separation and detection can be accomplished with a capillary-based electrophoresis platform commonly found in most forensic laboratories. © 2012 Springer Science+Business Media, LLC.