Comparison of Suitability of the Most Common Ancient DNA
Quantification Methods

J 2017

Comparison of Suitability of the Most Common Ancient DNA Quantification Methods

BRZOBOHATÁ, Kristýna, Eva DROZDOVÁ, Jiří SMUTNÝ, Tomáš ZEMAN, Radoslav BEŇUŠ et. al.

Základní údaje

Originální název

Comparison of Suitability of the Most Common Ancient DNA Quantification Methods

Autoři

BRZOBOHATÁ, Kristýna (203 Česká republika, garant, domácí), Eva DROZDOVÁ (203 Česká republika, domácí), Jiří SMUTNÝ (203 Česká republika), Tomáš ZEMAN (203 Česká republika) a Radoslav BEŇUŠ (703 Slovensko)

Vydání

Genetic Testing and Molecular Biomarkers, NEW ROCHELLE, USA, Mary Ann Liebert, Inc. 2017, 1945-0265

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

http://online.liebertpub.com/doi/abs/10.1089/gtmb.2016.0197

Impakt faktor

Impact factor: 1.181

Kód RIV

RIV/00216224:14310/17:00098659

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1089/gtmb.2016.0197

UT WoS

000398434800011

Klíčová slova anglicky

ancient DNA; aDNA quantification; PCR inhibition; aDNA fragmentation

Štítky

NZ, rivok

Změněno: 30. 3. 2018 15:32, Ing. Nicole Zrilić

Anotace

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.

Citovat

BRZOBOHATÁ, Kristýna, Eva DROZDOVÁ, Jiří SMUTNÝ, Tomáš ZEMAN a Radoslav BEŇUŠ. Comparison of Suitability of the Most Common Ancient DNA Quantification Methods. Genetic Testing and Molecular Biomarkers. NEW ROCHELLE, USA: Mary Ann Liebert, Inc., 2017, roč. 21, č. 4, s. 265-271. ISSN 1945-0265. Dostupné z: https://dx.doi.org/10.1089/gtmb.2016.0197.

@article{1398349,
   author = {Brzobohatá, Kristýna and Drozdová, Eva and Smutný, Jiří and Zeman, Tomáš and Beňuš, Radoslav},
   article_location = {NEW ROCHELLE, USA},
   article_number = {4},
   doi = {http://dx.doi.org/10.1089/gtmb.2016.0197},
   keywords = {ancient DNA; aDNA quantification; PCR inhibition; aDNA fragmentation},
   language = {eng},
   issn = {1945-0265},
   journal = {Genetic Testing and Molecular Biomarkers},
   title = {Comparison of Suitability of the Most Common Ancient DNA Quantification Methods},
   url = {http://online.liebertpub.com/doi/abs/10.1089/gtmb.2016.0197},
   volume = {21},
   year = {2017}
}

TY  - JOUR
ID  - 1398349
AU  - Brzobohatá, Kristýna - Drozdová, Eva - Smutný, Jiří - Zeman, Tomáš - Beňuš, Radoslav
PY  - 2017
TI  - Comparison of Suitability of the Most Common Ancient DNA Quantification Methods
JF  - Genetic Testing and Molecular Biomarkers
VL  - 21
IS  - 4
SP  - 265-271
EP  - 265-271
PB  - Mary Ann Liebert, Inc.
SN  - 19450265
KW  - ancient DNA
KW  - aDNA quantification
KW  - PCR inhibition
KW  - aDNA fragmentation
UR  - http://online.liebertpub.com/doi/abs/10.1089/gtmb.2016.0197
N2  - 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.
ER  -

BRZOBOHATÁ, Kristýna, Eva DROZDOVÁ, Jiří SMUTNÝ, Tomáš ZEMAN a Radoslav BEŇUŠ. Comparison of Suitability of the Most Common Ancient DNA Quantification Methods. \textit{Genetic Testing and Molecular Biomarkers}. NEW ROCHELLE, USA: Mary Ann Liebert, Inc., 2017, roč.~21, č.~4, s.~265-271. ISSN~1945-0265. Dostupné z: https://dx.doi.org/10.1089/gtmb.2016.0197.

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