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Updated: Jul 2, 2026

Measuring Single-Cell Mitochondrial DNA Copy Number and Heteroplasmy Using Digital Droplet Polymerase Chain Reaction
Published on: July 12, 2022
Sequencing strategy of mitochondrial HV1 and HV2 DNA with length heteroplasmy
E M Rasmussen1, E Sørensen, B Eriksen
1Department of Forensic Genetics, Institute of Forensic Medicine, University of Copenhagen, Frederik V's Vej 11, DK-2100 Copenhagen, Denmark. michael.rasmussen@forensic.ku.dk
Abstract:
We describe a method to obtain reliable mitochondrial DNA (mtDNA) sequences downstream of the homopolymeric stretches with length heteroplasmy in the sequencing direction. The method is based on the use of junction primers that bind to a part of the homopolymeric stretch and the first 2-4 bases downstream of the homopolymeric region. This junction primer method gave clear and unambiguous results using samples from 21 individuals with length heteroplasmy in the hypervariable regions HV1, HV2 or both. The method is of special value for forensic casework, because sequencing of both strands of an mtDNA region is preferable in order to reduce ambiguities in sequence determination.
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