摘要
为了探索一种适用于大豆叶片基因组DNA的快速、高效的提取方法,在传统的SDS提取方法的基础上,通过将缓冲液成分和浓度进行改良并加入表面活性剂Tween-20,优化实验流程。用限制性内切酶对该方法提取的DNA进行酶切及电泳检测,可得均一弥散条带。以提取的DNA模版扩增大豆Actin基因,测序后证实片段正确。同样以其为模版以SSR引物Sctt011进行SSR-PCR反应,并通过聚丙烯酰胺凝胶电泳检测,也可观察到特异性明显且无杂质的目的条带。实验结果表明,该方法提取的DNA完全符合各种分子实验的要求。本研究方法可用于快速提取大豆叶片基因组DNA,同时提高了提取DNA的质量。
In order to explore a method to fast extract high-quality soybean leaf genomic DNA.This study improved buffer composition and concentration,added surfactant Tween-20,and optimized the experimental procedure on the basis of the SDS method.The extracted DNA was digested with the restriction enzyme EcoR I,electrophoresis displayed it was digested completely.And the DNA was used as template to amplify the soybean Actin gene,sequencing confirmed the segment was correct.And the DNA was also used for SSR-PCR reaction with SSR primer Sctt011,polyacrylamide gel electrophoresis results showed the bands were specifical and clear.The results showed that the extracted DNA could be used for various molecular experiments.The method could shorten the DNA extraction time,while improve the quality of extracted DNA.
出处
《中国农学通报》
CSCD
2012年第9期38-41,共4页
Chinese Agricultural Science Bulletin
基金
国家科技部"863"计划重点项目"强优势大豆杂交种的创制与应用"(2011AA10A105)
国家转基因生物新品种培育科技重大专项"抗病虫转基因大豆新品种培育"(2011ZX08004-004)
吉林省青年科研基金项目"大豆细胞质雄性不育恢复基因的精细定位研究"(201201091)
关键词
大豆
基因组DNA
改良
快速
高质量
提取方法
soybean
genomic DNA
improvement
rapid
high-quality
extraction method
