蔓茎堇菜基因组DNA3种提取方法的比较研究

Comparison of Three Different Methods for the Isolation of Genomic DNA of Viol a diffusa Ging.

以SDS作为解离剂,采用常规SDS法、改进的SDS法及SDS高盐低pH法分离提取蔓茎堇菜基因组DNA.比较3种方法的提取效果,结果表明:常规SDS法DNA得率高但完整性及纯度较差;改进的SD S法D N A质量好但得率相对较低;只有SDS高盐低pH法是最为理想的提取方法,不同组织提取的DNA相对分子质量均大于48kb,260nm/280nm光密度比值在1.7～1.9之间,产率为479.0～543.9μg/g,所得DNA可直接用于限制性内切酶酶切,并适于进行RAPD分析.

Genomic DNA of Viola diffusa Ging. was isolated with three different methods using SDS as cleavage reagent, namely conventional SDS method, modified SDS method and low pH medium with high salts method （LPHS method）respectively. Results showed that conventional SDS method produced high DNA yield but the obtained genomic DNA was poor in terms of totality and purity. Modified SDS method produced good DNA in quality but low quantity in yield. LPHS method was the best and optimum way for DNA isolation in Viola diffusa Ging.. By this method, the length of the isolated DNA fragment from different tissues was over 48 kb and the 260 nm/280 nm of DNA was 1.7 - 1.9, the yield of the DNA was 479.0 - 543.9 μg/g and these DNA samples were all suitable for digestion with restriction enzyme and RAPD analysis.