芝麻油DNA高效提取方法的建立与优化

DEVELOPMENT AND OPTIMIZATION OF AN EFFICIENT EXTRACTION METHOD OF DNA FROM SESAME OIL

从植物油中提取高纯度DNA可为利用分子生物学技术检测油脂掺伪提供技术支撑。以芝麻油为材料优化了植物油DNA树脂提取法。在单因素试验的基础上,进行五因素四水平正交试验设计,以分光光度法测定提取DNA的浓度和纯度。得到芝麻油DNA提取的最优条件为:油样用量4 m L,去油液为4 m L氯仿,裂解液为2.5 m L 1.4 mol/L NaCl,摇匀时间50 min,摇匀速度25 r/min。在此条件下,DNA提取效果最佳。通过方差分析可知,摇匀速度对提取DNA浓度影响显著。

The molecular biology technology, PCR technology as the representative, has a good application prospect in edible oil adulteration detection and identification. Extraction of high purity DNA from vegetable oil is critical for the application of this method. The present study was to develop and optimize an efficient extraction method of DNA from sesame oil, while the orthogonal experiment of five factors and four levels was used on the basis of single-factor experiment. The DNA was isolated using spectrophotometric method, and the concentration and purity of DNA were determined by fluorescence spectrophotometry. The results showed that the optimal extraction conditions was as follows： oil sample amount of 4 mL, degreasing buffer of chloroform of 4 mL and lysis buffer of sodium chloride of 2.5 mL, the mixing time and mixing speed of 50 min and 25 r/min, respectively. ANOVA analysis was also performed and the results showed that the mixing rate had significant effect on the DNA concentration.