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Quality of DNA Extracted from Mouthwashes

BACKGROUND: A cost effective, safe and efficient method of obtaining DNA samples is essential in large scale genetic analyses. Buccal cells are an attractive source of DNA, as their collection is non-invasive and can be carried out by mail. However, little attention has been given to the quality of...

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Autores principales: Zayats, Tetyana, Young, Terri L., Mackey, David A., Malecaze, François, Calvas, Patrick, Guggenheim, Jeremy A.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2701599/
https://www.ncbi.nlm.nih.gov/pubmed/19582144
http://dx.doi.org/10.1371/journal.pone.0006165
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author Zayats, Tetyana
Young, Terri L.
Mackey, David A.
Malecaze, François
Calvas, Patrick
Guggenheim, Jeremy A.
author_facet Zayats, Tetyana
Young, Terri L.
Mackey, David A.
Malecaze, François
Calvas, Patrick
Guggenheim, Jeremy A.
author_sort Zayats, Tetyana
collection PubMed
description BACKGROUND: A cost effective, safe and efficient method of obtaining DNA samples is essential in large scale genetic analyses. Buccal cells are an attractive source of DNA, as their collection is non-invasive and can be carried out by mail. However, little attention has been given to the quality of DNA extracted from mouthwashes. METHODOLOGY: Mouthwash-derived DNA was extracted from 500 subjects participating in a genetic study of high myopia. DNA quality was investigated using two standard techniques: agarose gel electrophoresis and quantitative polymerase chain reaction (qPCR). PRINCIPAL FINDINGS: Whereas the majority of mouthwash-derived DNA samples showed a single band of high molecular weight DNA by gel electrophoresis, 8.9% (95% CI: 7.1–10.7%) of samples contained only a smear of low-to-medium molecular weight, degraded DNA. The odds of DNA degradation in a subject's second mouthwash sample, given degradation of the first, was significantly greater than one (OR = 3.13; 95% CI: 1.22–7.39; Fisher's test P = 0.009), suggesting that DNA degradation was at least partially a subject-specific phenomenon. Approximately 12.4% (95% CI: 10.4–14.4%) of mouthwash-derived DNA failed to PCR amplify efficiently (using an ∼200 bp microsatellite marker). However, we found there was no significant difference in amplification success rate between DNA samples judged to be degraded or non-degraded by gel electrophoresis (Fisher's test P = 0.5). CONCLUSIONS: This study demonstrated that DNA degradation affects a significant minority of saline mouthwashes, and that the phenomenon is partially subject-specific. Whilst the level of degradation did not significantly prevent successful amplification of short PCR fragments, previous studies suggest that such DNA degradation would compromise more demanding applications.
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spelling pubmed-27015992009-07-07 Quality of DNA Extracted from Mouthwashes Zayats, Tetyana Young, Terri L. Mackey, David A. Malecaze, François Calvas, Patrick Guggenheim, Jeremy A. PLoS One Research Article BACKGROUND: A cost effective, safe and efficient method of obtaining DNA samples is essential in large scale genetic analyses. Buccal cells are an attractive source of DNA, as their collection is non-invasive and can be carried out by mail. However, little attention has been given to the quality of DNA extracted from mouthwashes. METHODOLOGY: Mouthwash-derived DNA was extracted from 500 subjects participating in a genetic study of high myopia. DNA quality was investigated using two standard techniques: agarose gel electrophoresis and quantitative polymerase chain reaction (qPCR). PRINCIPAL FINDINGS: Whereas the majority of mouthwash-derived DNA samples showed a single band of high molecular weight DNA by gel electrophoresis, 8.9% (95% CI: 7.1–10.7%) of samples contained only a smear of low-to-medium molecular weight, degraded DNA. The odds of DNA degradation in a subject's second mouthwash sample, given degradation of the first, was significantly greater than one (OR = 3.13; 95% CI: 1.22–7.39; Fisher's test P = 0.009), suggesting that DNA degradation was at least partially a subject-specific phenomenon. Approximately 12.4% (95% CI: 10.4–14.4%) of mouthwash-derived DNA failed to PCR amplify efficiently (using an ∼200 bp microsatellite marker). However, we found there was no significant difference in amplification success rate between DNA samples judged to be degraded or non-degraded by gel electrophoresis (Fisher's test P = 0.5). CONCLUSIONS: This study demonstrated that DNA degradation affects a significant minority of saline mouthwashes, and that the phenomenon is partially subject-specific. Whilst the level of degradation did not significantly prevent successful amplification of short PCR fragments, previous studies suggest that such DNA degradation would compromise more demanding applications. Public Library of Science 2009-07-07 /pmc/articles/PMC2701599/ /pubmed/19582144 http://dx.doi.org/10.1371/journal.pone.0006165 Text en Zayats et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Zayats, Tetyana
Young, Terri L.
Mackey, David A.
Malecaze, François
Calvas, Patrick
Guggenheim, Jeremy A.
Quality of DNA Extracted from Mouthwashes
title Quality of DNA Extracted from Mouthwashes
title_full Quality of DNA Extracted from Mouthwashes
title_fullStr Quality of DNA Extracted from Mouthwashes
title_full_unstemmed Quality of DNA Extracted from Mouthwashes
title_short Quality of DNA Extracted from Mouthwashes
title_sort quality of dna extracted from mouthwashes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2701599/
https://www.ncbi.nlm.nih.gov/pubmed/19582144
http://dx.doi.org/10.1371/journal.pone.0006165
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