Detection of Heteroplasmic Variants in the Mitochondrial Genome through Massive Parallel Sequencing.

ABSTRACT: Detecting heteroplasmies in the mitochondrial DNA (mtDNA) has been a challenge for many years. In the past, Sanger sequencing was the main option to perform this analysis, however, this method could not detect low frequency heteroplasmies. Massive Parallel Sequencing (MPS) provides the opportunity to study the mtDNA in depth, but a controlled pipeline is necessary to reliably retrieve and quantify the low frequency variants. It has been shown that differences in methods can significantly affect the number and frequency of the retrieved variants. In this protocol, we present a method involving both wet lab and bioinformatics that allows identifying and quantifying single nucleotide variants in the full mtDNA sequence, down to a heteroplasmic load of 1.5%. For this, we set up a PCR-based amp

SUBMITTER: Mertens J 

PROVIDER: S-EPMC7854219 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature