超纯水对限制性内切酶酶切反应的影响

Influence of ultrapure water on restriction endonuclease digestion

目的 研究超纯水对限制性内切酶酶切结果的影响。方法 酶切体系中分别使用久置的超纯水和现用现配制的超纯水,以含酶切位点BamHⅠ和EcorⅤ的YFP-pcDNA3.1质粒为实验对象,双酶切后凝胶成像系统检测DNA琼脂糖凝胶电泳后的酶切结果。结果 应用久置的超纯水酶切结果有明显弥散现象,而更换现配制的超纯水,未见酶切弥散现象。结论 超纯水在分子生物学酶切实验中扮演重要角色,酶切实验应选用新制备的超纯水。

Objective To study the effects of different ultrapure water on the restriction endonuclease cleavage. Methods Ultrapure water placed in 4 ℃ refrigerator for a long time and fresh ultrapure water were applied in experiments respectively.Plasmid of YFP-pcDNA3.1 with BamHⅠ and Ecor Ⅴ enzyme cleavage sites was selected as the experimental subject.The results were acquired by gel imaging system after agarose gel electrophoresis. Results Significant diffusion phenomenon was observed when ultrapure water was applied which was placed in 4 ℃ refrigerator for a long time.however,no obvious diffusion phenomenon was observed when fresh ultrapure water was applied. Conclusion Fresh ultrapure water is essential for the experiment of restriction endonuclease cleavage.