An allelic ladder based upon reference alleles for mtDNA SNP analysis using the SNaPshot technique
✍ Scribed by Stephan Köhnemann; Petra Pennekamp; Heidi Pfeiffer
- Publisher
- John Wiley and Sons
- Year
- 2011
- Tongue
- English
- Weight
- 218 KB
- Volume
- 32
- Category
- Article
- ISSN
- 0173-0835
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
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.