Izvorni znanstveni članak
Development of a Multiplex Single Base Extension Assay for mitochondrial DNA Haplogroup Typing
Tahnee M. Nelson
Rebecca S. Just
Odile Loreille
Moses S. Schanfield
Daniele Podini
Sažetak
Aim To provide a screening tool to reduce time and sample consumption when attempting mitochondrial DNA (mtDNA) haplogroup
typing. Methods A single base primer extension assay was developed to enable typing, in a single reaction, of twelve mtDNA haplogroup specific polymorphisms. For validation purposes a total of 147 samples were tested including 73 samples successfully haplogroup typed using mtDNA control region (CR) sequence data, 20 samples inconclusively haplogroup typed by CR sequence data, 21 samples previously haplogroup typed using restriction fragment length polymorphism (RFLP) analysis, and 31 samples of known ancestral origin without previous haplogroup typing. Additionally, two highly degraded human bones embalmed and buried in the early 1950s were analyzed using the single nucleotide polymorphisms (SNP) multiplex. Results When the SNP multiplex was used to type the 96 previously CR sequenced specimens, an increase in haplogroup or macrohaplogroup assignment relative to conventional CR sequence analysis was observed. The single base extension assay was also successfully used to assign a haplogroup to decades-old, embalmed skeletal remains dating to World War II. Conclusion The SNP multiplex was successfully used to obtain haplogroup status of highly degraded human bones, and demonstrated the ability to eliminate possible contributors. The SNP multiplex provides a low-cost, high throughput method for typing of mtDNA haplogroups A, B, C, D, E, F, G, H, L1/L2, L3, M, and N that could be useful for screening purposes for human identification efforts and anthropological studies.
Ključne riječi
mitochondrial DNA (mtDNA); bones; haplogroups
Hrčak ID:
16585
URI
Datum izdavanja:
15.8.2007.
Posjeta: 2.123 *