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An allelic ladder based upon reference alleles for mtDNA SNP analysis using the SNaPshot technique
Author(s) -
Köhnemann Stephan,
Pennekamp Petra,
Pfeiffer Heidi
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201100124
Subject(s) - genetics , biology , single nucleotide polymorphism , allele , snapshot (computer storage) , snp , mitochondrial dna , multiplex , snp genotyping , computational biology , genotype , gene , computer science , operating system
The analysis of mitochondrial DNA (mtDNA) single‐nucleotide polymorphisms (SNPs) using the SNaPshot technique (Applied Biosystems) is a fast and sensitive method for the reliable identification of disease‐associated mtDNA SNPs, genetic ancestry mtDNA SNPs and forensically important mtDNA SNPs. The detection of many SNPs in one multiplex PCR and one subsequent multiplex minisequencing reaction is challenging for laboratories who want to establish this technique, due to the problem that there is no allelic ladder available for mtDNA SNP analysis via SNaPshot technique. Normally, the laboratory has to invent long‐term testing and studies. The interpretation of false and correct alleles is up to some specialists knowing the expected and the estimated size of each allele SNP. We here present a protocol to assemble up to 84 alleles of 42 different mtDNA SNPs in an allelic ladder that is based upon reference alleles. We recommend using allelic ladders/reference alleles for SNP analysis to maintain high‐quality analysis standards.