食用植物油DNA提取方法的优化

Optimization of DNA extraction from edible vegetable oil

中文关键词:  食品安全  食用植物油  DNA提取  十六烷基三甲基溴化铵法  实时荧光定量PCR

英文关键词:food safety  edible vegetable oil  DNA extraction  cetyltrimethylammonium bromide method  quantitative real-time PCR

中文摘要:

      目的 建立可从不同种类食用植物油中提取得到高质量的、可用于分子生物学检测的DNA的提取方法。方法 使用2种改进十六烷基三甲基溴化铵法(cetyltrimethylammonium bromide, CTAB)和2种商用试剂盒提取6种食用植物油的DNA, 通过紫外分光光度计检测所得DNA样品的浓度和纯度。设计特异性引物, 并通过普通聚合酶链式反应(polymerase chain reaction, PCR)和实时荧光定量PCR(quantitative real-time PCR, qPCR)检测DNA样品是否可用于分子生物学分析。结果 使用改良CTAB法1及2种商业试剂盒无法有效提取得到的6种食用植物油的DNA, 样品无法满足分子生物学检测的需要, 使用改良CTAB法2提取得到的6种食用植物油DNA纯度和浓度检测结果均为最佳, 其提取的橄榄油、菜籽油、花生油DNA样品可用于后续实时荧光定量PCR检测。结论 该方法可为橄榄油、菜籽油、花生油DNA提取提供有效手段, 为食用植物油检测和研究奠定基础。

英文摘要:

      Objective To establish a DNA extraction method which can extract high-quality DNA from different types of edible vegetable oils and can be used for molecular biological testing. Methods Two modified cetyltrimethylammonium bromide (CTAB) methods and 2 commercial kits were used to extract the DNA of 6 edible vegetable oils. The concentration and purity of the obtained DNA samples were detected by an ultraviolet spectrophotometer. Specific primers were designed, and common polymerase chain reaction (PCR) and quantitative real-time PCR (q-PCR) were used to detect whether DNA samples could be used for molecular biological analysis. Results The DNA of 6 edible vegetable oils could not be effectively extracted by the modified CTAB method 1 and 2 commercial kits, and the samples could not meet the needs of molecular biology testing. While the modified CTAB method 2 could be used to extract DNA from 6 edible vegetable oils of which the concentration and purity were superior in quality. The extracted DNA samples from olive oil, colza oil, and peanut oil could be used for q-PCR detection, respectively. Conclusion This study provides an effective method for olive oil, colza oil, and peanut oil DNA extraction, and lays a foundation for edible vegetable oil detection and research.