[Real-time quantification to analyze historical Colombian samples detecting a short fragment of hypervariable region II of mitochondrial DNA].
Biomedica
; 36(3): 475-482, 2016 Sep 01.
Article
em Es
| MEDLINE
| ID: mdl-27869396
ABSTRACT
INTRODUCTION:
Unlike other molecular biology studies, the analysis of ancient DNA (aDNA) requires special infrastructure and methodological conditions to guarantee the quality of the results. One of the main authenticity criteria is DNA quantification, where quantitative real-time PCR is often used given its sensitivity and specificity. Nevertheless, the implementation of these conditions and methodologies to fulfill authenticity criteria imply higher costs.Objective:
To develop a simple and less costly method for mitochondrial DNA quantification suitable for highly degraded samples. Materials andmethods:
The proposed method is based on the use of mini-primers for the specific amplification of short fragments of mitochondrial DNA. The subsequent purification of these amplified fragments allows a standard curve to be constructed with concentrations in accordance to the state of degradation of the samples.Results:
The proposed method successfully detected DNA from ancient samples including bone remains and mummified tissue. DNA inhibitory substances were also detected.Conclusion:
The proposed method represents a simpler and cost-effective way to detect low amounts of aDNA, and a tool to differentiate DNA-free samples from samples with inhibitory substances.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Mitocondrial
/
Reação em Cadeia da Polimerase em Tempo Real
Limite:
Humans
País/Região como assunto:
America do sul
/
Colombia
Idioma:
Es
Revista:
Biomedica
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
Colômbia