【摘要】 目的:比较提取高质量艰难梭菌基因组总DNA的方法。方法:采用4种不同方法提取艰难梭菌基因组DNA,比较作为模板用于扩增艰难梭菌看家基因磷酸丙糖异构酶(tpi)基因的优缺点。结果:除沸水浴法,其他3种方法制备的艰难梭菌DNA均能成功用作PCR扩增的模板。CTAB/NaCl法进行提取可有效去除多糖成分,但操作要求较高。碱裂解法操作简便、省时、成本低经济可靠,提取的DNA量及纯度较合适;试剂盒抽提出的产物纯度和产量明显高于CTAB/NACL、碱裂解法(P<0.05),但成本高。结论:四种方法各具优缺点,应根据实验室条件和实验要求进行选择。更多还原

【Abstract】 Objective: To compare the relative efficiency and quality of extraction of C.difficile DNA using four different methods.Methods: Extract C.difficile genomic DNA with four different methods: immersion method,CTAB degrading solution,Alkali solit method and kit.To amplified the special gene(tpi) of C.difficile by real-time PCR.Results: The products by the three methods are suitable for real-time PCR amplification,but except immersion method.Extraction with CTAB degrading solution could remove the polysaccharide,but the method demands more operating skill.Alkali solit method offered a higher OD260/280(1.80±0.06) and this method is simpler and cheaper.The purity and output of DNA extracted by kit were obviously higher(P<0.05),but the method was more amplicated and expensive.Conclusion: The four methods all have advantages and disadvantages of their own,and which method to use should be based on the lab conditions and research demands.更多还原