双酶切反应通用缓冲液使用表

DoubleDigestion（双酶切反应）时UniversalBuffer（通用缓冲液）的使用■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UniversalBuffer之前表示的［数字习是指各UniversalBuffer的反应体系中的最终浓度。TaKaRa销售产品中添附的UniversalBuffer全为10倍浓度的缓冲液。终浓度为0.5用寸反应体系中的缓冲液则稀释至20倍，1X0寸稀释至10倍，2X时稀释至5倍进行使用。■注息◊1卬gDNA中添加10U的限制酶，在50"勺反应体系中，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。EnzymeAccIBamHiBglWC!a\EcoR1EcoRVHineIIH/ndHIKpn1Neo1NdelAcc1IxM0.5xKIxT1xM1xM0.5xK1xM1xM1xMIxMfBSAIxTfia/nH10.5XK1XKIxK1xK1xK1XK0.5XK1xK0.5XKixK+BSAIxKBglII1XT1XK1XH1xH1XH1XH2XK1xK1XTIxKiBSA1xHClalIxM1XK1XH1xM1XH1xH1XM1XM1xM1XK+BSA1xH£coRIIxM1xK1XH1XH1XH1xH1xM1XM1xMIxKfBSA1xHfcoRV0.5xKIxK1XH1xH1XH1xH2xT1XK0.5XK1XK+BSA1xHHineII1xM0.5xK2xK1XM1xM2xT1xM1xM1xM1乂瞄SA1xTHindHlIxM1xKIXK1xM1xM1xK1xM1xM1xMixKtBSAIxKKpn1IxM05xK1XT1XM1xM0.5xK1xM1XM1XL0.5XK48SA1xTNeo1IxM+BSA1xK+BSAixK+BSAIxK曲IxK-hBSAIxIGBSAIxM^BSAIxK^BSA0.5xK+BSAIxK+BSA1XKtBSANdel1XT1XK1XH1XH1xH1XH1XT1XK1XT1XK+8SA1xHNotl0.5xKfBSA0.5XK-BSAixHiBSAIxHiBSAixH^BSAIXH^BSA05XK.BSA05XK.BSAOSXK+BSA0.5XK+8SA1xH.BSAPstl1xM1xK1XH1XH1XH1XH1XM1XM1xMIxK+BSA1xHPvuI0.5xKIxK1xK1XK1XK1xK0.5KK1xK0.5XKIxK+BSAIxKSac11XM05XK0.5XK1XM1XM0.5XK1XM1XM1XLOixKf&SA1xTSai\1.5x11.5XT1xH1XH1XH1XH1.5XK1.5XK15XT+BSA15XT+BSA1xHSma1ixhBSA0.5XKBSA1XT+BSA1XT4BSA1XT4BSAOSxK+BSA1xT+BSA1XKBSAixT+BSA1XT+BSAIxT^BSASpe1IxMIxKIxH1xM1xH1xH1xM1xM1xMIxK+BSA1xHSph10.5xK1xK1xH1XH1xH1xH2xT1xK0.5XK1XK.8SA1xHXb3\IxM03xK2xT1XM1XM2xT1xM1xM1XM1M8SA1xTXho\IxM1xK1xH1xH1xH1xH1xM1xM1xM1xK1xHEnzymeNotIPstlPvu\Sac\Sa/1Sma1SpelSph1Xba\Xho\Acc\0.5xKtBSA1xM0.5xK1xM1.5xT1xTtBSA1xM0.5xK1xM1xMBamHI0.5XK成A1xK1xK0.5xK1.5XT0.5XT+BSA1xKIxK0.5XK1xKBgl\l1XH+ESAIxH1xK0.5xK1xH1xT+BSA1xHIxH2xT1xHCla\1XHfBSA1xH1xK1xM1乂H1xT+BSA1xM1xH1乂M1xHEcoR11XH+BSA1XH1xK1xM1xH1xT+BSA1xH1xH1xM1xH£"RVIxH+BSA1XH1xK0.5xK1XH05XK+BSA1xH1XH2xT1XHHineII0.5XK+8SA1xM0.5XK1xM1.5XK1xT+BSA1xM2XT1XM1xMHind1110.5xKtB$A1xM1xK1xM1.5xK1xTtBSA1xMIxK1xM1xMKpnl05XK+BSA1XM0.5XK1XLLSxT+BSA1XT+BSA1xM0.5xK1XM1xMNeoI05xK*8SA1XK+B5A1XK+BSA0.5xKtBSA1.5xT+BSAIxTtBSA1xK+BSA1XK+BSA1xM+BSA1xK+BSANde\1XH+ESA1xH1xK1XT1xH1xT+BSA1xH1xH1XT1xHNotIixHfCSA+1rtonX-1001XH+BSA2xK+BSA0.5xKf8SA1XH+BSA0.5XT+BSA1XH+BSA1XWBSA0.5xK#BSA1XH4BSAPsfl1xH+ESAIxH1xK1xM1xH0.5xTtBSA1xH1xH1xM1xHPvuI2XK+ESA1xKIXK+BSA0.5xK1.5XK4BSA1xK+BSA1xKIXK0.5XK1xKSacI0.5xKfBSA1xM0.5xK1xL1时般1xT+BSA1xM0.5xK1xM1xMSail1XH+BSA1xHISxKfBSA1.5XT+BSA1xH1.5XT+BSA1xH1xH1.5XT1xHSmaI0.5XT4SA0.5XT+BSA1XK*BSA1XT+BSA1WSA1XT+BSAIxT^BSA0.5XUBSAixT+BSA1XT+BSASpeIIxH+ESAIxH1xK1xM1xH1xTtBSA1xM1xH1xM1xHSphI1XH+ESA1XH1xK0.5XK1xH0.5XT+BSA1XH1XH2xT1xHXbal0.5XK/&SA1XM0.5XK1xM1.5XT1xT+BSA1xM2xT1XM+BSA1xMXho\1xH1xH1xK1xM1xH1XT1xH1xH1XM1xH