摘要
为筛选出用于大豆DNA提取的适宜方法,分别使用Bayer法、DuPont法、Monsanto法、Qiagen和Tiangen试剂盒法提取大豆子粒DNA,并通过紫外分光光度法、琼脂糖凝胶电泳法与实时荧光聚合酶链式反应分析DNA质量。结果显示:Bayer法提取的DNA含有较多杂质,抑制实时荧光聚合酶链式反应;Monsanto法提取的DNA纯度差且得率低,不符合检测要求;DuPont法、Qiagen与Tiangen试剂盒法能提取到纯度好、得率高、完整性好的DNA,符合实时荧光聚合酶链式反应的检测要求,但是Qiagen试剂盒法成本过高。因此,DuPont法与Tiangen试剂盒法适用于大豆转基因检测中的DNA提取。
The objective of this study was to screen a suitable method for DNA extraction from soybean by comparison of five generally applied DNA extraction methods. After extraction, the quality of soybean DNA was examined with ultraviolet spectrophotometry, agarose gel electrophoresis and real-time fluorescent PCR, respectively. The results showed that the DNA extracted by Bayer method contained many PCR inhibitors. The purity and yield of DNA extracted by Monsanto method did not meet the demand of quality standard of DNA in molecular experiment. DuPont method, Qiagen kit and Tiangen kit could extract high-quality DNA which was suitable for real-time fluorescent PCR, but Qiagen kit cost too much. These evidences indicated that DuPont method and Tiangen kit were suitable methods to extract DNA from soybean for detection of genetically modified organism （GMO）.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第4期199-203,共5页
Food Science
基金
转基因生物新品种培育重大专项(2009ZX08012-014B)
江苏省疾病预防控制中心青年基金项目(JSQ200511)
关键词
大豆
DNA提取
转基因生物
琼脂糖凝胶电泳
实时荧光聚合酶链式反应
soybean
DNA extraction
genetically modified organism
agarose gel electrophoresis
real-time polymerase chain reaction （PCR）
