利用改良CTAB法提取淮山叶片高质量DNA

Extraction of High Quality DNA from Leaves of Dioscorea oposetap Thunb.with Modified CTAB Method

[目的]为淮山的分子生物学研究奠定基础。[方法]以淮山嫩叶为试材,用改良CTAB法提取其中的DNA,并通过琼脂糖凝胶电泳和ISSR-PCR扩增对所得DNA的质量和浓度进行检测,考察该方法对DNA的提取效果。[结果]所提取DNA的分子量与λDNA相近,各泳道均出现1条清晰的DNA条带,且条带的完整性较好,无蛋白质和RNA污染,无DNA降解现象;以所提DNA为模板进行ISSR-PCR扩增,可获得稳定的扩增条带。[结论]改良CTAB法简单高效,所得DNA质量较高,RNA消化彻底,DNA无明显降解,进行ISSR分析时,条带清晰,多态性较好。

[ Objective] The study was to provide the basis for molecular biology research on Dioscorea oposetap Thunb. [ Method] With young leaves of D. oposetap as the materials, their DNA was extracted with modified CTAB method. The quality and concn, of the extracted DNA were detected by agarose gel eIectrophoresis and ISSR-PCR amplification. The extracting effect of modified CTAB method on DNA was stud- ied. [ Result] The molecular weight of the extracted DNA was similar with that of kDNA. There was 1 clear DNA strip in each lane,and the integrity of the strip was better without protein and RNA pollution and DNA degradation phenomenon. The stable amplified strips could be obtained through ISSR-PCR amplification with template of the extracted DNA. [ Conclusion ] The modified CTAB method was simple with high effficient,and the quality of DNA extracted by this method was higher,the digestion of RNA was complete,and there were no obvious degradation phenomenon of DNA, and the strip was clear with better polymorphism in ISSR analysis.