Rapid Screening for Native American Mitochondrial and Y-Chromosome

GALA ZUCCARELLI GALA; EVGUENIA ALECHINE; MARIELA CAPUTO; CECILIA BOBILLO; ANDREA SALA

FORENSIC SCIENCE INTERNATIONAL-GENETICS

ELSEVIER IRELAND LTD

Lugar: Ireland; Año: 2010

1872-4973

Aiming to detect individuals of Native American maternal or paternal ancestry a rapid screening approach has been developed. Its strategy was based on SNP typing by Real Time PCR (rt-PCR) followed by High Resolution Melting analysis (HRM). After extraction, DNA was quantitated by rt-PCR using commercial kits; samples were then submitted to two multiplex reactions in order to determine the major Native American mtDNA and Y-chromosome haplogroups by HRM. One cocktail included primers flanking nucleotide substitutions that define mtDNA haplogroup C and sub-haplogroups A2, B2, and D1. The other included primers flanking Y-SNPs M3, M269 and U179 that allowed discriminating Q and non-Q hgs. In all cases amplicons were less than 125 nucleotides long in order to increase the peak resolution. The accuracy of the results obtained was established by means of sequencing analysis of the amplicons. The new working-flow here proposed facilitates and speeds-up the screening process that may preclude a detailed sequencing analysis of particular samples, or for further molecular epidemiological investigations in which continental origin influences might be relevant.