Nedel Fernanda, Conde Marcus Cristian Muniz, Oliveira Isabel Oliveira de, Tarquinio Sandra Beatriz Chaves, Demarco Flávio Fernando
Dental School, Federal University of Pelotas, Pelotas, RS, Brazil.
Braz Dent J. 2009;20(4):275-8. doi: 10.1590/s0103-64402009000400002.
This study compared quantitatively and qualitatively the DNA extracted from buccal cells collected from the upper or lower gutter areas. Buccal cells were collected from the upper (n=15) and lower gutter (n=15) region from 15 volunteers using a special cytobrush (Gentra), totaling 2 collections from each individual. DNA was extracted from the samples according to the manufacturer's instructions. The DNA obtained was qualitatively and quantitatively evaluated by 2 calibrated blind examiners using spectrophotometry and analysis of DNA bands (0.8% agarose gel electrophoresis). Data was statistically analyzed by one-way ANOVA (alpha=0.05). Means and standard derivation (SD) for total DNA yield from the upper and lower gutter area were 12.2 microg (12.0) and 9.4 microg (8.5), respectively (p=0.821). There was higher (p<0.05) DNA purity for the upper gutter (1.79; 0.05) when compared to lower gutter area (1.66; 0.10). Regarding to the DNA quality, no differences were observed between the 2 location sites, but all samples showed similar degree of degradation. In conclusion, it would be recommendable that buccal cells for DNA extraction be collected from the upper gutter area in the attempt to increase DNA purity.
本研究对从上颌沟或下颌沟区域采集的颊细胞中提取的DNA进行了定量和定性比较。使用特殊的细胞刷（Gentra）从15名志愿者的上颌沟（n = 15）和下颌沟（n = 15）区域采集颊细胞，每人共采集2份样本。按照制造商的说明从样本中提取DNA。由2名经过校准的盲法检测人员使用分光光度法和DNA条带分析（0.8%琼脂糖凝胶电泳）对获得的DNA进行定性和定量评估。数据采用单因素方差分析进行统计学分析（α = 0.05）。上颌沟和下颌沟区域总DNA产量的平均值和标准差（SD）分别为12.2微克（12.0）和9.4微克（8.5）（p = 0.821）。与下颌沟区域（1.66；0.10）相比，上颌沟的DNA纯度更高（p < 0.05）（1.79；0.05）。关于DNA质量，在两个采集部位之间未观察到差异，但所有样本均显示出相似程度的降解。总之，为提高DNA纯度，建议从上颌沟区域采集用于DNA提取的颊细胞。
