西南桦硅胶干燥叶DNA提取方法比较

Comparison on DNA Extraction Methods from Betula alnoides Leaves Dried with Silica Gel

采用常规CTAB法、CTAB硅珠法、核分离法、二次抽提法、改良CTAB法对富含多糖的西南桦干叶进行DNA提取,并将DNA进行电泳检测及RAPD分析,结果表明:不同提取方法结果差异较大,CTAB硅珠法得率最低,为0.4～11.6μg/g;核分离法和二次抽提法较常规法去除多糖效果好,得率高;改良CTAB法得率最高,为171.2～364.8μg/g,电泳条带最清晰,RAPD扩增结果最好,适合干叶西南桦基因组DNA提取。

The total genomic DNA was extracted from the dried leaves of Betula alnoides with CTAB,CTAB-SiO2,the nuclear separation,double times extraction,and modified CTAB methods,and the DNA obtained was analyzed by the means of agarose gel electrophoresis and RAPD analysis.The results showed that there was great difference in DNA yield from different extraction methods.The DNA yield by CTAB-SiO2 method was the lowest with 0.4-11.6 μg/g.The DNA yield obtained by the nuclear separation and double times extraction methods was higher than the CTAB method,which could eliminate the glucose more efficiently.The DNA yield from the modified CTAB method was the highest,which was 171.2-364.8 μg / g,the polymorphic patterns amplified by RAPD were the best with clear electrophoresis belts.It was concluded that the modified CTAB method was suitable to extract the total genomic DNA from dried leaves of Betula alnoides.