Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains

Ancient DNA (aDNA) recovered from archaeobotanical remains can provide key insights into many prominent archaeological research questions, including processes of domestication, past subsistence strategies, and human interactions with the environment. However, it is often difficult to isolate aDNA fr...

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Main Authors: Wales, N., Andersen, K., Cappellini, E., Avila-Arcos, M., Gilbert, Thomas
Format: Journal Article
Published: Public Library of Science 2014
Online Access:http://hdl.handle.net/20.500.11937/23723
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author Wales, N.
Andersen, K.
Cappellini, E.
Avila-Arcos, M.
Gilbert, Thomas
author_facet Wales, N.
Andersen, K.
Cappellini, E.
Avila-Arcos, M.
Gilbert, Thomas
author_sort Wales, N.
building Curtin Institutional Repository
collection Online Access
description Ancient DNA (aDNA) recovered from archaeobotanical remains can provide key insights into many prominent archaeological research questions, including processes of domestication, past subsistence strategies, and human interactions with the environment. However, it is often difficult to isolate aDNA from ancient plant materials, and furthermore, such DNA extracts frequently contain inhibitory substances that preclude successful PCR amplification. In the age of high-throughput sequencing, this problem is even more significant because each additional endogenous aDNA molecule improves analytical resolution. Therefore, in this paper, we compare a variety of DNA extraction techniques on primarily desiccated archaeobotanical remains and identify which method consistently yields the greatest amount of purified DNA. In addition, we test five DNA polymerases to determine how well they replicate DNA extracted from non-charred ancient plant remains. Based upon the criteria of resistance to enzymatic inhibition, behavior in quantitative real-time PCR, replication fidelity, and compatibility with aDNA damage, we conclude these polymerases have nuanced properties, requiring researchers to make educated decisions as to which one to use for a given task. The experimental findings should prove useful to the aDNA and archaeological communities by guiding future research methodologies and ensuring precious archaeobotanical remains are studied in optimal ways, and may thereby yield important new perspectives on the interactions between humans and past plant communities.
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spelling curtin-20.500.11937-237232017-09-13T14:01:06Z Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains Wales, N. Andersen, K. Cappellini, E. Avila-Arcos, M. Gilbert, Thomas Ancient DNA (aDNA) recovered from archaeobotanical remains can provide key insights into many prominent archaeological research questions, including processes of domestication, past subsistence strategies, and human interactions with the environment. However, it is often difficult to isolate aDNA from ancient plant materials, and furthermore, such DNA extracts frequently contain inhibitory substances that preclude successful PCR amplification. In the age of high-throughput sequencing, this problem is even more significant because each additional endogenous aDNA molecule improves analytical resolution. Therefore, in this paper, we compare a variety of DNA extraction techniques on primarily desiccated archaeobotanical remains and identify which method consistently yields the greatest amount of purified DNA. In addition, we test five DNA polymerases to determine how well they replicate DNA extracted from non-charred ancient plant remains. Based upon the criteria of resistance to enzymatic inhibition, behavior in quantitative real-time PCR, replication fidelity, and compatibility with aDNA damage, we conclude these polymerases have nuanced properties, requiring researchers to make educated decisions as to which one to use for a given task. The experimental findings should prove useful to the aDNA and archaeological communities by guiding future research methodologies and ensuring precious archaeobotanical remains are studied in optimal ways, and may thereby yield important new perspectives on the interactions between humans and past plant communities. 2014 Journal Article http://hdl.handle.net/20.500.11937/23723 10.1371/journal.pone.0086827 Public Library of Science fulltext
spellingShingle Wales, N.
Andersen, K.
Cappellini, E.
Avila-Arcos, M.
Gilbert, Thomas
Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains
title Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains
title_full Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains
title_fullStr Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains
title_full_unstemmed Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains
title_short Optimization of DNA recovery and amplification from non-carbonized archaeobotanical remains
title_sort optimization of dna recovery and amplification from non-carbonized archaeobotanical remains
url http://hdl.handle.net/20.500.11937/23723