一种经济高效的用Silica提取纯化质粒DNA的方法  

A Rapid and High Efficiency Method for Plasmid DNA Extraction and Purification Using Silica

作　　者：周国雁[1] 郭凤根[1] 张应华[1] 杨正安[1] 孙茂林[2] 丁玉梅[2] 

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]云南省农业生物技术重点实验室,云南昆明650223

出　　处：《生物技术通讯》2007年第5期800-802,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(C02021001);云南省自然科学基金项目(2004C0025Q)

摘　　要：目的:为了达到批量提取质粒DNA的目的,在多次实验的基础上,建立一种经济、高效的质粒提取方法。方法:以pUC18、pET28b、pCAMBIA1304等3种质粒为材料,分别采用silica法和碱裂解法提取质粒DNA,通过质粒DNA浓度的紫外分光光度法定量测定、电泳分析和Hind Ⅲ酶切鉴定,对两种质粒提取方法的效果进行了比较与评价;对silica法进行了改进和优化,进行大批量重组子的提取和验证。结果:silica法和碱裂解法提取质粒DNA效果相当,都可进行后续实验,但silica法具有经济、高效、无毒的优势。结论:silica法是一种简单、经济、高效的质粒提取方法,可用于批量质粒DNA提取。Objective： To get an efficient method for batches of plasmid DNA extraction, a simple, inexpensive but effective method based on many times of experiments was developed. Methods： Two methods, alkaline lysis and silica, have been used for extraction of plasmid DNA from pUC18, pET28b and pCAMBIA1304. By quantitative analysis of DNA concentration, agarose electrophoresis and restriction enzyme digestion of Hind Ⅲ, the effects of two plasmid DNA extraction methods were evaluated. Finally, the method of silica was improved and optimized. Results： The effects of two methods were quite the same, plasmid DNA can be used to the following experiments. However, using silica for plasmid DNA isolation had the superiority of cheapness, high-efficiency and innocuity. Conclusion： The results indicated that silica can be able to applied in batch extraction of plasmid DNA, it is a more efficient, time and money-saving method for plasmid DNA isolation.

关 键 词：质粒DNA 提取 碱裂解法 SILICA 

分 类 号：Q503[生物学—生物化学]