takara_双酶切buffer用表.doc

Enzyme Acc I BamH I Bgl II Cla I EcoR I EcoR V Hinc II Hind III Kpn I Nco I Nde I Not I Pst I Pvu I Sac I Sal I Sma I Spe I Sph I Xba I Xho ISupplied Buffer 10 ×M 10 ×K 10 ×H 10 ×M 10 ×H 10 ×H 10 ×M 10 ×M 10 ×L 10 ×K+BSA 10 ×H10 ×H+BSA +Triton10 ×H 10 ×K+BSA 10 ×L 10 ×H 10 ×T+BSA 10 ×M 10 ×H 10 ×M+BSA 10 ×HAcc I - 0.5 × K 1 ×T 1 ×M 1 ×M 0.5 × K 1 ×M 1 ×M 1 ×M 1 ×M+BSA 1 ×T 0.5 × K+BSA 1 ×M 0.5 × K 1 ×M 1.5 × T 1 ×T+BSA 1 ×M 0.5 × K 1 ×M 1 ×MBamH I 0.5 × K - 1 ×K 1 ×K 1 ×K 1 ×K 0.5 × K 1 ×K 0.5 × K 1 ×K+BSA 1 ×K 0.5 × K+BSA 1 ×K 1 ×K 0.5 × K 1.5 × T 0.5 × T+BSA 1 × K 1 × K 0.5 × K 1 × KBgl II 1 × T 1 × K - 1 × H 1 × H 1 × H 2 × K 1 × K 1 × T 1 × K+BSA 1 × H 1 × H+BSA 1 × H 1 × K 0.5 × K 1 × H 1 × T+BSA 1 × H 1 × H 2 × T 1 × HCla I 1 × M 1 × K 1 × H - 1 × H 1 × H 1 × M 1 × M 1 × M 1 × K+BSA 1 × H 1 × H+BSA 1 × H 1 × K 1 × M 1 × H 1 × T+BSA 1 × M 1 × H 1 × M 1 × HEcoR I 1 × M 1 × K 1 × H 1 × H - 1 × H 1 × M 1 × M 1 × M 1 × K+BSA 1 × H 1 × H+BSA 1 × H 1 × K 1 × M 1 × H 1 × T+BSA 1 × H 1 × H 1 × M 1 × HEcoR V 0.5 × K 1 × K 1 × H 1 × H 1 × H - 2 × T 1 × K 0.5 × K 1 × K+BSA 1 × H 1 × H+BSA 1 × H 1 × K 0.5 × K 1 × H 0.5 × K+BSA 1 × H 1 × H 2 × T 1 × HHinc II 1 × M 0.5 × K 2 × K 1 × M 1 × M 2 × T - 1 × M 1 × M 1 × M+BSA 1 × T 0.5 × K+BSA 1 × M 0.5 × K 1 × M 1.5 × K 1 × T+BSA 1 × M 2 × T 1 × M 1 × MHind III 1 × M 1 × K 1 × K 1 × M 1 × M 1 × K 1 × M - 1 × M 1 × K+BSA 1 × K 0.5 × K+BSA 1 × M 1 × K 1 × M 1.5 × K 1 × T+BSA 1 × M 1 × K 1 × M 1 × MKpn I 1 × M 0.5 × K 1 × T 1 × M 1 × M 0.5 × K 1 × M 1 × M - 0.5 × K+BSA 1 × T 0.5 × K+BSA 1 × M 0.5 × K 1 × L 1.5 × T+BSA 1 × T+BSA 1 × M 0.5 × K 1 × M 1 × MNco I 1 × M+BSA 1 × K+BSA 1 × K+BSA 1 × K+BSA 1 × K+BSA 1 × K+BSA 1 × M+BSA 1 × K+BSA 0.5 × K+BSA - 1 × K+BSA 0.5 × K+BSA 1 × K+BSA 1 × K+BSA 0.5 × K+BSA 1.5 × T+BSA 1 × T+BSA 1 × K+BSA 1 × K+BSA 1 × M+BSA 1 × K+BSANde I 1 × T 1 × K 1 × H 1 × H 1 × H 1 × H 1 × T 1 × K 1 × T 1 × K+BSA - 1 × H+BSA 1 × H 1 × K 1 × T 1 × H 1 × T+BSA 1 × H 1 × H 1 × T 1 × HNot I 0.5 × K+BSA 0.5 × K+BSA 1 × H+BSA 1 × H+BSA 1 × H+BSA 1 × H+BSA 0.5 × K+BSA 0.5 × K+BSA 0.5 × K+BSA 0.5 × K+BSA 1 × H+BSA - 1 × H+BSA 2 × K+BSA 0.5 × K+BSA 1 × H+BSA 0.5 × T+BSA 1 × H+BSA 1 × H+BSA 0.5 × K+BSA 1 × H+BSAPst I 1 × M 1 × K 1 × H 1 × H 1 × H 1 × H 1 × M 1 × M 1 × M 1 × K+BSA 1 × H 1 × H+BSA - 1 × K 1 × M 1 × H 0.5 × T+BSA 1 × H 1 × H 1 × M 1 × HPvu I 0.5 × K 1 × K 1 × K 1 × K 1 × K 1 × K 0.5 × K 1 × K 0.5 × K 1 × K+BSA 1 × K 2 × K+BSA 1 × K - 0.5 × K 1.5 × K+BSA 1 × K+BSA 1 × K 1 × K 0.5 × K 1 × KSac I 1 × M 0.5 × K 0.5 × K 1 × M 1 × M 0.5 × K 1 × M 1 × M 1 × L 0.5 × K+BSA 1 × T 0.5 × K+BSA 1 × M 0.5 × K - 1.5 × T+BSA 1 × T+BSA 1 × M 0.5 × K 1 × M 1 × MSal I 1.5 × T 1.5 × T 1 × H 1 × H 1 × H 1 × H 1.5 × K 1.5 × K 1.5 × T+BSA 1.5 × T+BSA 1 × H 1 × H+BSA 1 × H 1.5 × K+BSA 1.5 × T+BSA - 1.5 × T+BSA 1 × H 1 × H 1.5 × T 1 × HSma I 1 × T 0.5 × T 1 × T 1 × T 1 × T 0.5 × K 1 × T 1 × T 1 × T 1 × T 1 × T 0.5 × T 0.5 × T 1 × K 1 × T 1.5 × T - 1 × T 0.5 × T 1 × T 1 × T+BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSA +BSASpe I 1 × M 1 × K 1 × H 1 × M 1 × H 1 × H 1 × M 1 × M 1 × M 1 × K+BSA 1 × H 1 × H+BSA 1 × H 1 × K 1 × M 1 × H 1 × T+BSA - 1 × H 1 × M 1 × HSph I 0.5 × K 1 × K 1 × H 1 × H 1 × H 1 × H 2 × T 1 × K 0.5 × K 1 × K+BSA 1 × H 1 × H+BSA 1 × H 1 × K 0.5 × K 1 × H 0.5 × T+BSA 1 × H - 2 × T 1 × HXba I 1 × M 0.5 × K 2 × T 1 × M 1 × M 2 × T 1 × M 1 × M 1 × M 1 × M+BSA 1 × T 0.5 × K+BSA 1 × M 0.5 × K 1 × M 1.5 × T 1 × T+BSA 1 × M 2 × T - 1 × MXho I 1 × M 1 × K 1 × H 1 × H 1 × H 1 × H 1 × M 1 × M 1 × M 1 × K+BSA 1 × H 1 × H+BSA 1 × H 1 × K 1 × M 1 × H 1 × T+BSA 1 × H 1 × H 1 × M -Note: 1) It is confirmed that 10 units of each enzyme completely digest 1 µg of DNA at 37°C in one hour in 50 µl reaction mixture. 2) The concentration of Glycerol should be less than 10% to minimize star activity. 3) DNA may not be digested completely, when recognition sequences of two enzymes are close each other, or when DNA takes high-structure conformation.Relative activity for each reaction buffer: supplied, activity measured buffer: recommended bufferRelative activities (%)Restriction enzymeL M H K T+BSA Basal***Aat II 1/10 volume of 10× Loading Buffer to stop enzyme reaction and apply on agarose gel electrophoresis. SDS may precipitate during the storage at room temperature. In case precipitates generate, dissolve in warm bath before use.。