AustralieFrV1, Main, Exploration, bibRecord, 007B67

Quality of DNA Extracted from Mouthwashes

Identifieur interne : 007B67 ( Main/Exploration ); précédent : 007B66; suivant : 007B68

Quality of DNA Extracted from Mouthwashes

Auteurs : Tetyana Zayats [Royaume-Uni] ; Terri L. Young [États-Unis] ; David A. Mackey [Australie] ; François Malecaze [France] ; Patrick Calvas [France] ; Jeremy A. Guggenheim [Royaume-Uni]

Source :

PLoS ONE [ 1932-6203 ] ; 2009.

RBID : PMC:2701599

Abstract

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.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2701599

DOI: 10.1371/journal.pone.0006165
PubMed: 19582144
PubMed Central: 2701599

Affiliations:

Le document en format XML

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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>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.</p>
</sec>
<sec><title>Methodology</title>
<p>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).</p>
</sec>
<sec><title>Principal Findings</title>
<p>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).</p>
</sec>
<sec><title>Conclusions</title>
<p>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.</p>
</sec>
</div>
</front>
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<author><name sortKey="Le Marchand, L" uniqKey="Le Marchand L">L Le Marchand</name>
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<author><name sortKey="Bu, Dw" uniqKey="Bu D">DW Bu</name>
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<author><name sortKey="Lewis, C" uniqKey="Lewis C">C Lewis</name>
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<affiliations><list><country><li>Australie</li>
<li>France</li>
<li>Royaume-Uni</li>
<li>États-Unis</li>
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<region><li>Caroline du Nord</li>
<li>Midi-Pyrénées</li>
<li>Occitanie (région administrative)</li>
<li>Victoria (État)</li>
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<settlement><li>Melbourne</li>
<li>Toulouse</li>
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<orgName><li>Université de Melbourne</li>
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<tree><country name="Royaume-Uni"><noRegion><name sortKey="Zayats, Tetyana" sort="Zayats, Tetyana" uniqKey="Zayats T" first="Tetyana" last="Zayats">Tetyana Zayats</name>
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<country name="États-Unis"><region name="Caroline du Nord"><name sortKey="Young, Terri L" sort="Young, Terri L" uniqKey="Young T" first="Terri L." last="Young">Terri L. Young</name>
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<country name="Australie"><region name="Victoria (État)"><name sortKey="Mackey, David A" sort="Mackey, David A" uniqKey="Mackey D" first="David A." last="Mackey">David A. Mackey</name>
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<name sortKey="Mackey, David A" sort="Mackey, David A" uniqKey="Mackey D" first="David A." last="Mackey">David A. Mackey</name>
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<country name="France"><region name="Occitanie (région administrative)"><name sortKey="Malecaze, Francois" sort="Malecaze, Francois" uniqKey="Malecaze F" first="François" last="Malecaze">François Malecaze</name>
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<name sortKey="Calvas, Patrick" sort="Calvas, Patrick" uniqKey="Calvas P" first="Patrick" last="Calvas">Patrick Calvas</name>
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   |texte=   Quality of DNA Extracted from Mouthwashes
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Serveur d'exploration sur les relations entre la France et l'Australie

Quality of DNA Extracted from Mouthwashes

Quality of DNA Extracted from Mouthwashes

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Abstract

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	Serveur d'exploration sur les relations entre la France et l'Australie
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