【摘要】 对提取植物DNA的CTAB法进行了改进。使用Retsch研磨仪极大地提高了效率,并在CTAB提取液中添加0.2%的二硫苏糖醇可以有效防止酚类物质的氧化及与DNA的不可逆结合。紫外吸收检测DNAA260/A280=1.7￣1.8;经琼脂糖凝胶电泳检测得到一条带型较宽且清晰的DNA谱带,辣椒叶片DNA平均产量为29.8μg/100mg,证明所提取DNA的质量和得率均较高;酶切结果表明DNA质量较高,能够酶切完全;PCR扩增到的特异性条带明亮、整齐,基本无拖尾,达到各种分子标记等研究技术的要求。更多还原

【Abstract】 There are many of amylose and hydroxybenzene in pepper leaves. It is difficult to extract high quality DNA by formal methods including method CTAB, which is very fussy for grinding by liquid nitrogen. We optimized the extraction methods of CTAB and Grinded pepper leaves by Retsch to increase the extraction efficiency greatly. The addition of 0.1% DTT into CTAB extraction reagent can prevent the hydroxybenzene from oxygenation and decrease the hydroxybenzene to bind with DNA. DNA extracted by this method had a A260/A280=1.7-1.8 detected by ultraviolet radiation absorbance and got a wider and bright band in the gelose electrophoresis. The average output of DNA is 29.8 μg DNA/100 mg pepper leaves. DNA extracted by this method has a high purity and output. The results of enzyme incubation and PCR with these DNA as templates show that they are good to molecular marker analysis and other biology research.更多还原