Detailed Information on Publication Record
2017
Comparison of Suitability of the Most Common Ancient DNA Quantification Methods
BRZOBOHATÁ, Kristýna, Eva DROZDOVÁ, Jiří SMUTNÝ, Tomáš ZEMAN, Radoslav BEŇUŠ et. al.Basic information
Original name
Comparison of Suitability of the Most Common Ancient DNA Quantification Methods
Authors
BRZOBOHATÁ, Kristýna (203 Czech Republic, guarantor, belonging to the institution), Eva DROZDOVÁ (203 Czech Republic, belonging to the institution), Jiří SMUTNÝ (203 Czech Republic), Tomáš ZEMAN (203 Czech Republic) and Radoslav BEŇUŠ (703 Slovakia)
Edition
Genetic Testing and Molecular Biomarkers, NEW ROCHELLE, USA, Mary Ann Liebert, Inc. 2017, 1945-0265
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10608 Biochemistry and molecular biology
Country of publisher
United Kingdom of Great Britain and Northern Ireland
Confidentiality degree
není předmětem státního či obchodního tajemství
Impact factor
Impact factor: 1.181
RIV identification code
RIV/00216224:14310/17:00098659
Organization unit
Faculty of Science
UT WoS
000398434800011
Keywords in English
ancient DNA; aDNA quantification; PCR inhibition; aDNA fragmentation
Změněno: 30/3/2018 15:32, Ing. Nicole Zrilić
Abstract
V originále
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.