CAJ | 학술논문

高质量的DNA是进行基因克隆等分子生物学试验的前提条件.因大豆种子中蛋白质和油脂含量丰富,利用传统方法提取DNA质量很难达到试验要求.研究在SDS提取液的基础上.通过添加表面活性剂NP-40和Tween-20,优化出一种适合于大豆种子DNA快速提取的方法.优化后的SDS方法提取的DNA质量较高.OD_(260)/OD_(280)在1.809～1.916之间,无蛋白质和RNA污染.对该方法提取的DNA进行了基因克隆和分子标记的试验验证,结果表明,它们能够用于大豆基因克隆、分子标记.
고질량적DNA시진행기인극륭등분자생물학시험적전제조건.인대두충자중단백질화유지함량봉부,이용전통방법제취DNA질량흔난체도시험요구.연구재SDS제취액적기출상.통과첨가표면활성제NP-40화Tween-20,우화출일충괄합우대두충자DNA쾌속제취적방법.우화후적SDS방법제취적DNA질량교고.OD_(260)/OD_(280)재1.809～1.916지간,무단백질화RNA오염.대해방법제취적DNA진행료기인극륭화분자표기적시험험증,결과표명,타문능구용우대두기인극륭、분자표기.
High-quality DNA is an important base of molecular biology manipulation, such as gene cloning. There are lots of protein and fat in the soybean seeds, so the DNA extracted from them by using traditional methods is very difficult to achieve quality requirements of experiments. This study improved the SDS extraction buffer by adding NP-40 and Tween-20, and established a rapid DNA extraction method that is suitable for soybean seeds. The DNA extraction by the optimized SDS extraction method has high-quality, and the values of OD_(260)/OD_(280) ranged from 1.809 to 1.916, that means non-protein and RNA pollution. The DNA extraction method was validated by gene cloning and genotyping, and the results showed that it could be used for soybean gene cloning and molecular markers.