摘要
目的:比较不同DNA提取方法,选择最适于中麻黄基因组的方法。方法:采用改良CTAB,SDS法、试剂盒法提取中麻黄基因组DNA;用琼脂糖凝胶电泳法和紫外分光光度法检测所得总DNA的得率和纯度,并进行ISSR-PCR扩增检测。结果:3种方法均可从新鲜中麻黄中提取出产量较高的基因组DNA,但其中改良CTAB法提取的总DNA纯度高,质量好,扩增产物的电泳条带也较明显;SDS法和试剂盒法提取的总DNA质量差,不适用于下游分子生物学实验。结论:改良十六烷基三甲基溴化铵(CTAB)法为中麻黄基因组DNA提取的最佳方法,该方法提取的基因组DNA适用于中麻黄基因组PCR扩增和其他分子生物学研究。
Objective： By comparing different extraction methods of genomic DNA, the best extraction method of Ephedra intermedia genomic DNA was selected. Method： Improved cetyhrimethyl ammonium bromide （CTAB） method, SDS method, kit method were used for the extraction of genomic DNA from E. intermedia. The yield and purity of total genomic DNA were examined by agarose gel electrophoresis, UV-spectrophotometry and ISSR-PCR. Result： Three extraction methods could extracted the high amount genomic DNA from fresh tissue of E. intermedia. Improved CTAB method could yield highest purity total DNA, and the electrophoretic bands of amplified products were obvious. On the other hand, SDS method and kit method indicated low purity of the yielded DNA and could not be used in downstream application directly. Conclusion： The improved CTAB method is the best of the three compared methods for extracting the genomic DNA from E. intermedia, which is suitable for PCR reaction and other molecular research in E. intermedia genomic DNA.
出处
《中国实验方剂学杂志》
CAS
北大核心
2012年第11期125-128,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
科技基础性工作专项重点项目子课题(SB2007FY020)
甘肃教育厅科研项目(1106B-07)
甘肃中医学院中青年科研基金项目(ZQ2011-12)
作者简介
朱田田，硕士，讲师，从事中药资源开发与利用研究，Tel：13088731903，E-mail：ztt0935@163．com
[通讯作者]晋玲，博士，副教授，从事药用植物资源可持续利用研究，Tel：0931-8765335，E-mail：jinl@gszy．edu．cn
