DoubleDigestion(双酶切反应)时UniversalBuffer(通用缓冲液)的使用表

1、DoubleDigestion（双酶切反应）时UniversalBuffer（通用缓冲液）的使用表说明使用二种酶同时进行DNA切断反应（DoubleDigestion）时，为了节省反应时间，通常希望在同一反应体系内进行。TaKaRa采用UniversalBuffer表示系统，并对每种酶表示了在各UniversalBuffer中的相对活性。尽管如此，在进行DoubleDigestion时，有时还会难以找到合适的UniversalBuffer。本表以在pUC系列载体的多克隆位点处的各限制酶为核心，显示了在DoubleDigestion可使用的最佳UniversalBuffer条件。在本表中，各Un

2、iversalBuffer之前表示的数字x是指各UniversalBuffer的反应体系中的最终浓度oTaKaRa销售产品中添附的UniversalBuffer全为10倍浓度的缓冲液。终浓度为0.5x时反应体系中的缓冲液则稀释至20倍，1x时稀释至10倍,2x时稀释至5倍进行使用。注意1pgDNA中添加10U的限制酶，在50pl的反应体系中，37C下反应1小时可以完全降解DNA。为防止Star活性的产生，请将反应体系中的甘油含量，尽量控制在10%以下。根据DNA的种类，各DNA的立体结构的差别，或当限制酶识别位点邻接时，有时会发生DoubleDigestion不能顺利进行的可能。Nde1xT1

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