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从漱口水提取的DNA质量。

Quality of DNA extracted from mouthwashes.

作者信息

Zayats Tetyana, Young Terri L, Mackey David A, Malecaze François, Calvas Patrick, Guggenheim Jeremy A

机构信息

School of Optometry and Vision Sciences, Cardiff University, Wales, UK.

出版信息

PLoS One. 2009 Jul 7;4(7):e6165. doi: 10.1371/journal.pone.0006165.

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 approximately 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.

摘要

背景

在大规模基因分析中,一种经济高效、安全且有效的获取DNA样本的方法至关重要。颊细胞是一种有吸引力的DNA来源,因为其采集是非侵入性的,并且可以通过邮寄进行。然而,对于从漱口液中提取的DNA质量关注较少。

方法

从500名参与高度近视基因研究的受试者中提取漱口液来源的DNA。使用两种标准技术研究DNA质量:琼脂糖凝胶电泳和定量聚合酶链反应(qPCR)。

主要发现

虽然大多数漱口液来源的DNA样本通过凝胶电泳显示出一条高分子量DNA条带,但8.9%(95%置信区间:7.1 - 10.7%)的样本仅含有低至中等分子量的降解DNA条带。如果第一个样本发生DNA降解,那么受试者第二个漱口液样本发生DNA降解的几率显著大于1(比值比 = 3.13;95%置信区间:1.22 - 7.39;费舍尔检验P = 0.009),这表明DNA降解至少部分是个体特异性现象。大约12.4%(95%置信区间:10.4 - 14.4%)的漱口液来源的DNA未能有效进行PCR扩增(使用约200 bp微卫星标记)。然而,我们发现通过凝胶电泳判断为降解或未降解的DNA样本之间的扩增成功率没有显著差异(费舍尔检验P = 0.5)。

结论

本研究表明,DNA降解影响了相当一部分盐水漱口液样本,且该现象部分是个体特异性的。虽然降解程度并未显著妨碍短PCR片段的成功扩增,但先前的研究表明,这种DNA降解会影响更具挑战性的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5be3/2701599/a186ac528a73/pone.0006165.g001.jpg

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