限制性内切酶反应是分子生物学实验中的一个重要方法。基因物理图谱的绘制、DNA核苷酸序列分析、基因片段的体外切割重组及转化菌中重组子的筛选等等,都需要使用限制性内切酶。因此,限制性内切酶反应做的如何也是关键的一步。鼠疫菌质粒DNA的限制性内切酶反应的实验结果如下。1 材料与方法 菌株 由本室提供鼠疫强毒株。 限制性内切酶 EcoRV(带缓冲液),中国协和医科大学医学科学技术开发公司。 鼠疫菌质粒DNA提取用碱裂解法。 限制性内切酶反应 参照有关文献并略加改动。 取鼠疫菌质粒DNA8μl,依次加入10μl蒸馏水、2μl10×缓冲液和1～2单位限制性内切酶。37℃水浴1～2小时,加0.4μl0.5mol/L EDTA(pH8.0)终止 Restriction enzyme reaction is an important method in molecular biology experiments. Gene mapping of the physical map, DNA nucleotide sequence analysis, in vitro cutting and recombination of gene fragments and selection of transformants recombinant recombinants, etc., require the use of restriction enzymes. Therefore, how the restriction enzyme reaction is done is also a crucial step. The experimental results of the restriction endonuclease reaction of the plague plasmid DNA are as follows. 1 Materials and methods Strain from the room to provide plague virulent strains. Restriction enzyme EcoRV (with buffer), Peking Union Medical College Medical Science and Technology Development Company. Yersinia pestis plasmid DNA extraction by alkaline lysis method. Restriction enzyme reaction with reference to the literature and slightly modified. Take plague plasmid DNA 8μl, followed by the addition of 10μl distilled water, 2μl 10 × buffer and 1 ~ 2 units of restriction endonuclease. 37 ℃ water bath for 1 to 2 hours, add 0.4μl0.5mol / L EDTA (pH8.0) termination