质粒抽提策略对双酶切鉴定的影响

Effect of the strategy of extracting plasmids on the double digestion identification

目的探讨质粒抽提策略对质粒双酶切鉴定的影响。方法比较1.4 ml菌液单次抽提与分成两份0.7 ml和四份0.35 ml多次抽提的质粒双酶切后琼脂糖凝胶电泳图谱质量。结果小体积菌液多次抽提的质粒纯度逐步提高(p ET-28a和p ET-28a-G6PD的OD260/OD280值分别为1.36和1.45,1.65和1.70,1.75和1.78);酶切后琼脂糖凝胶电泳弥散逐步消失。结论控制菌液量对抽提高纯度质粒、获得理想的双酶切鉴定图谱至关重要。

Objective To explore the effect of the strategy of plasmid extraction on the double digestion identification. Methods Plasmids were extracted according to three stategies：1 × 1.4 ml bacterium solution for extraction,2 × 0.7 ml for extraction,4 × 0.35 ml for extraction. The image qualities of double digested plasmids displayed in agarose gel electrophoresis were compared. Results The smears gradually disappeared and the purities of plasmids gradually rose as the bacterium solution for plasmids extraction reduced. Conclusion Controlling the amount of bacterium is crucial for obtaining high purity of plasmids and ideal images of double digestion i- dentification.