珍稀濒危树种金钱槭的基因组DNA提取方法研究  

Study on Genomic DNA Extraction Methods for the Rare and Endangered Tree Species Dipteronia sinensis Oliv.

作　　者：吴晟昊 杨丽 向松竹 胡茜茜 周志翔[1,2] 郑波 施雪萍 刘梅[2] WU Sheng-hao;YANG Li;XIANG Song-zhu(College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Wuhan,Hubei 430070;Hubei Shennongjia Academy of Forestry,Shennongjia,Hubei 442400)

机构地区：[1]华中农业大学园艺林学学院,湖北武汉430070 [2]湖北神农架林区林业科学研究院,湖北神农架442400

出　　处：《安徽农业科学》2024年第3期91-94,共4页Journal of Anhui Agricultural Sciences

基　　金：神农架林区林业科学研究院科技计划项目“神农架林区珍稀濒危树种香果树和金钱槭繁育技术研究”(SAF202107)。

摘　　要：[目的]明确高效提取金钱槭基因组DNA的方法。[方法]以金钱槭幼嫩叶片为材料,采用4种不同方法提取金钱槭基因组DNA,并通过紫外分光光度计法、琼脂糖凝胶电泳检测、限制性内切酶酶切和扩增保守基因片段等方法比较不同DNA的提取效率。[结果]高盐低pH法因操作时间较长导致DNA损失较多,提取到的DNA虽然杂质含量较低,但浓度也较低。SDS法提取效果不佳,提取到的DNA含量少且完整性差。尿素法提取的DNA质量最差,降解严重,难以用于后续分子生物学研究。改良CTAB法可以大量提取能够用于酶切及PCR扩增的DNA。[结论]金钱槭DNA高效提取方法的确定可以为相关的分子生物学研究提供科学依据和技术参考。[Objective]To determine efficient method for extracting genomic DNA from D.sinensis.[Method]Four methods were used to extract genomic DNA from young leaves of D.sinensis.The efficiency of genomic DNA extraction was assessed by ultraviolet spectrophotometer,agarose gel electrophoresis,restriction enzyme digestion,and amplification of conserved gene fragments.[Result]The results showed that the high-salt,low-pH method led to more DNA loss due to the extended operation time,resulting in low concentration of DNA with low impurity content.The SDS method had a poor extraction efficiency,yielding DNA with low content and poor integrity.The urea method produced the worst DNA extraction,with severe degradation that renders it unsuitable for molecular biology research.The improved CTAB method successfully extracted a large amount of DNA suitable for enzyme digestion and PCR amplification.[Conclusion]The determination of an efficient DNA extraction method will provide scientific basis and technical references for related molecular biology research of D.sinensis.

关 键 词：金钱槭 DNA提取 CTAB法 

分 类 号：S792.35[农业科学—林木遗传育种]