Zayats, Tetyana, Young, T. L., Mackey, D. A., Malecaze, F., Calvas, P. and Guggenheim, Jeremy Andrew ORCID: https://orcid.org/0000-0001-5164-340X 2009. Quality of DNA extracted from mouthwashes. PLoS ONE 4 (7) , e6165. 10.1371/journal.pone.0006165 |
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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.
Item Type: | Article |
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Date Type: | Publication |
Status: | Published |
Schools: | Optometry and Vision Sciences |
Subjects: | Q Science > QH Natural history > QH426 Genetics |
Publisher: | Public Library of Science |
ISSN: | 1932-6203 |
Last Modified: | 09 May 2023 09:15 |
URI: | https://orca.cardiff.ac.uk/id/eprint/11325 |
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