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用SDS高盐介质法和1.5×CTAB法提取黑木相思幼叶及叶状柄基因组DNA,将其与2×CTAB法、3×CTAB法、改进的CTAB法进行比较,并用紫外光谱吸收、凝胶电泳、限制性内切酶消化、RAPD(Radomly Amplified Polymorphic DNA)和ISSR(Inter-Simple Sequence Repeat)等方法进行鉴定.结果表明,叶状柄多糖的质量分数较幼叶少,更易于基因组DNA的提取;SDS高盐介质法与1.5×CTAB法提取叶状柄的基因组DNA得率为180.0~697.5μg.g-1,分子质量为48 kb,D(260)/D(280)=1.750~1.955,无需经RNase处理,可直接用于限制性酶切和RAPD及ISSR分析;1.5×CTAB法所提的基因组DNA的纯度优于SDS高盐介质法.综合考虑认为,1.5×CTAB法可作为黑木相思基因组DNA快速提取的简便方法.
Genomic DNA was extracted from young leaves and leaf stems of Acacia meya using SDS high salt medium and 1.5 × CTAB method, compared with 2 × CTAB method, 3 × CTAB method and modified CTAB method, Gel electrophoresis, restriction endonuclease digestion, RAPD (Radomly Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat), etc. The results showed that the phyllostem polysaccharide had less mass fraction than the young leaves, The yield of genomic DNA extracted from leafy stems by SDS high salt medium and 1.5 × CTAB method was 180.0 ~ 697.5μg.g-1, the molecular mass was 48 kb, D (260) / D (280) = 1.750 ~ 1.955, without RNase treatment, can be directly used for restriction analysis and RAPD and ISSR analysis; 1.5 × CTAB method of genomic DNA purity is superior to the SDS high salt medium method. Comprehensive consideration that the 1.5 × CTAB method can be used as A Simple and Convenient Method for Quickly Extracting Genomic DNA from Acacia mangium.