International Journal of Legal Medicine

, Volume 122, Issue 5, pp 385–388

‘Mitominis’: multiplex PCR analysis of reduced size amplicons for compound sequence analysis of the entire mtDNA control region in highly degraded samples

Original Article

DOI: 10.1007/s00414-008-0227-5

Cite this article as:
Eichmann, C. & Parson, W. Int J Legal Med (2008) 122: 385. doi:10.1007/s00414-008-0227-5

Abstract

The traditional protocol for forensic mitochondrial DNA (mtDNA) analyses involves the amplification and sequencing of the two hypervariable segments HVS-I and HVS-II of the mtDNA control region. The primers usually span fragment sizes of 300–400 bp each region, which may result in weak or failed amplification in highly degraded samples. Here we introduce an improved and more stable approach using shortened amplicons in the fragment range between 144 and 237 bp. Ten such amplicons were required to produce overlapping fragments that cover the entire human mtDNA control region. These were co-amplified in two multiplex polymerase chain reactions and sequenced with the individual amplification primers. The primers were carefully selected to minimize binding on homoplasic and haplogroup-specific sites that would otherwise result in loss of amplification due to mis-priming. The multiplexes have successfully been applied to ancient and forensic samples such as bones and teeth that showed a high degree of degradation.

Keywords

mtDNA CR Mini-amplicons Multiplex Forensic science 

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Institute of Legal MedicineInnsbruck Medical UniversityInnsbruckAustria

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