Quality of DNA extracted from mouthwashes.
Quality of DNA extracted from mouthwashes.
复制标题
DOI：
10.1371/journal.pone.0006165
复制
发表时间：
2009-07-07
期刊：
影响因子：
3.7
通讯作者：
Guggenheim JA
中科院分区：
文献类型：
--
作者：
Zayats T;Young TL;Mackey DA;Malecaze F;Calvas P;Guggenheim JA
在大规模遗传分析中，一种经济、安全和高效的DNA样本获取方法是必不可少的。口腔细胞是一个有吸引力的DNA来源，因为它们的收集是非侵入性的，可以通过邮件进行。然而，从漱口水中提取的DNA的质量却鲜有人关注。从500名参与高度近视基因研究的受试者中提取了含漱口水的DNA。DNA质量检测采用两种标准技术：琼脂糖凝胶电泳法和定量聚合酶链式反应(QPCR)法。然而，大多数含漱口水的DNA样本在凝胶电泳中显示出单一的高分子量DNA条带，8.9%(95%可信区间：7.1-10.7%)的样本仅含有低至中等分子质量的降解DNA涂片。考虑到第一个样本的降解，受试者第二个漱口水样本中的脱氧核糖核酸降解的几率明显大于一个样本(OR = 3.13；95%CI：1.22-7.39；费舍尔检验P = 0.009)，这表明脱氧核糖核酸的降解至少部分是受试者特有的现象。大约12.4%(95%可信区间：10.4-14.4%)的含漱液中的dna未能有效地扩增(使用∼200bp微卫星标记)。但经凝胶电泳法判定为降解和未降解的样品，其扩增成功率无显著差异(Fisher‘s检验P = .5)。这项研究表明，DNA降解影响了相当少数的含盐漱口水，而且这种现象部分是特定于受试者的。虽然降解的程度并没有显著地阻止短的PCR片段的成功扩增，但以前的研究表明，这种DNA降解会损害更苛刻的应用。
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. 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). 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). 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.
登录
查看更多内容
影响因子：
3.9
作者：
Bergen, AW;Haque, KA;Chanock, SJ
通讯作者：
Chanock, SJ
影响因子：
4.4
作者：
Farbrother, JE;Kirov, G;Guggenheim, JA
通讯作者：
Guggenheim, JA
影响因子：
2.6
作者：
Glei, M;Habermann, N;Pool-Zobel, BL
通讯作者：
Pool-Zobel, BL
影响因子：
3.3
作者：
König, KG
通讯作者：
König, KG
影响因子：
6.7
作者：
Haffajee, AD;Socransky, SS
通讯作者：
Socransky, SS