Comparison of Suitability of the Most Common Ancient DNA Quantification Methods

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F17%3A00098659" target="_blank" >RIV/00216224:14310/17:00098659 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1089/gtmb.2016.0197" target="_blank" >http://dx.doi.org/10.1089/gtmb.2016.0197</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1089/gtmb.2016.0197" target="_blank" >10.1089/gtmb.2016.0197</a>

Result language

angličtina

Original language name

Comparison of Suitability of the Most Common Ancient DNA Quantification Methods

Original language description

Aims: Ancient DNA (aDNA) extracted from historical bones is damaged and fragmented into short segments, present in low quantity, and usually copurified with microbial DNA. A wide range of DNA quantification methods are available. The aim of this study was to compare the five most common DNA quantification methods for aDNA. Materials and Methods: Quantification methods were tested on DNA extracted from skeletal material originating from an early medieval burial site. The tested methods included ultraviolet (UV) absorbance, real-time quantitative polymerase chain reaction (qPCR) based on SYBR® green detection, real-time qPCR based on a forensic kit, quantification via fluorescent dyes bonded to DNA, and fragmentary analysis. Differences between groups were tested using a paired t-test. Results: Methods that measure total DNA present in the sample (NanoDrop™ UV spectrophotometer and Qubit® fluorometer) showed the highest concentrations. Methods based on real-time qPCR underestimated the quantity of aDNA. The most accurate method of aDNA quantification was fragmentary analysis, which also allows DNA quantification of the desired length and is not affected by PCR inhibitors. Conclusions: Methods based on the quantification of the total amount of DNA in samples are unsuitable for ancient samples as they overestimate the amount of DNA presumably due to the presence of microbial DNA. Real-time qPCR methods give undervalued results due to DNA damage and the presence of PCR inhibitors. DNA quantification methods based on fragment analysis show not only the quantity of DNA but also fragment length.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

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Continuities

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Genetic Testing and Molecular Biomarkers

ISSN

1945-0265

e-ISSN

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Volume of the periodical

21

Issue of the periodical within the volume

4

Country of publishing house

GB - UNITED KINGDOM

Number of pages

7

Pages from-to

265-271

UT code for WoS article

000398434800011

EID of the result in the Scopus database

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