桫椤科植物DNA提取方法研究  

Extraction Methods of Genomic DNA from Cyatheaceae Plant

作　　者：应站明[1] 应正河[2] 潘伟华[3] 

机构地区：[1]萍乡高等专科学校,江西萍乡337000 [2]福建省农科院食用菌研究所,福建福州350014 [3]中山大学生命科学学院,广东广州510275

出　　处：《安徽农业科学》2010年第8期3922-3924,共3页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(30771763)

摘　　要：[目的]探讨桫椤科植物DNA提取的最优方法。[方法]采用经过改良的SDS法和CTAB法对3种桫椤科植物进行基因组DNA提取,用紫外分光光度计测定其在波长260和280nm处的吸光值,并根据A260nm/A280nm的比值分析其DNA的纯度。将提取的基因组DNA进行叶绿体trnL-trnF非编码区序列扩增和ISSR分析。[结果]改进的CTAB法所提出的DNA较纯净,所获得DNA样品的A260nm/A280nm比值在1.750～2.050范围内,琼脂糖凝胶电泳图谱显示条带清晰。[结论]改良的CTAB法更适合桫椤科植物DNA的提取。[Objective] To explore an efficient method for extracting genomic DNA from Cyatheaceae plants.[Method] The improved sodium dodecy sulphate （SDS） and cety triethl ammonium bromide （CTAB） methods were used for the genomic DNA extraction of three kinds of Cyatheaceae plants.The absorption values were deter mined by ultraviolet spectrophotometry at A260 nm and A280 nm,and the DNA purities were analyzed by A260 nm/A280 nm ratio.The amplification of chloroplast trnL-trnF non-coding region and ISSR analysis of genomic DNA were then conducted.[Result] The results showed that the improved CTAB method of extracting genomic DNA from Cyatheaceae plants was better than the improved SDS method,and the DNA purity of the former was higher than that of the latter.The A260 nm/A280 nm ratio of the extracted DNA with the improved CTAB method was in the range from 1.750 to 2.050,and distinct bands were found in the agarose gel electrophoresis map of ISSR.[Conclusion] The improved CTAB method is more suitable for the DNA extraction of Cyatheaceae plant.

关 键 词：桫椤科 CTAB法 SDS法 ISSR 

分 类 号：S312[农业科学—作物栽培与耕作技术]