Takara 双酶切buffer用表.docx

1、Takara 双酶切buffer用表EnzymeAcc IBamH IBgl IICla IEcoR IEcoR VHinc IIHind IIIKpn INco INde INot IPst IPvu ISac ISal ISma ISpe ISph IXba IXho ISupplied Buffer10 M10 K10 H10 M10 H10 H10 M10 M10 L10 K+BSA10 H10 H+BSA +Triton10 H10 K+BSA10 L10 H10 T+BSA10 M10 H10 M+BSA10 HAcc I-0.5 K1 T1 M1 M0.5 K1 M1 M1 M1

2、 M+BSA1 T0.5 K+BSA1 M0.5 K1 M1.5 T1 T+BSA1 M0.5 K1 M1 MBamH I0.5 K-1 K1 K1 K1 K0.5 K1 K0.5 K1 K+BSA1 K0.5 K+BSA1 K1 K0.5 K1.5 T0.5 T+BSA1 K1 K0.5 K1 KBgl II1 T1 K-1 H1 H1 H2 K1 K1 T1 K+BSA1 H1 H+BSA1 H1 K0.5 K1 H1 T+BSA1 H1 H2 T1 HCla I1 M1 K1 H-1 H1 H1 M1 M1 M1 K+BSA1 H1 H+BSA1 H1 K1 M1 H1 T+BSA1 M

3、1 H1 M1 HEcoR I1 M1 K1 H1 H-1 H1 M1 M1 M1 K+BSA1 H1 H+BSA1 H1 K1 M1 H1 T+BSA1 H1 H1 M1 HEcoR V0.5 K1 K1 H1 H1 H-2 T1 K0.5 K1 K+BSA1 H1 H+BSA1 H1 K0.5 K1 H0.5 K+BSA1 H1 H2 T1 HHinc II1 M0.5 K2 K1 M1 M2 T-1 M1 M1 M+BSA1 T0.5 K+BSA1 M0.5 K1 M1.5 K1 T+BSA1 M2 T1 M1 MHind III1 M1 K1 K1 M1 M1 K1 M-1 M1 K+

4、BSA1 K0.5 K+BSA1 M1 K1 M1.5 K1 T+BSA1 M1 K1 M1 MKpn I1 M0.5 K1 T1 M1 M0.5 K1 M1 M-0.5 K+BSA1 T0.5 K+BSA1 M0.5 K1 L1.5 T+BSA1 T+BSA1 M0.5 K1 M1 MNco I1 M+BSA1 K+BSA1 K+BSA1 K+BSA1 K+BSA1 K+BSA1 M+BSA1 K+BSA0.5 K+BSA-1 K+BSA0.5 K+BSA1 K+BSA1 K+BSA0.5 K+BSA1.5 T+BSA1 T+BSA1 K+BSA1 K+BSA1 M+BSA1 K+BSANd

5、e I1 T1 K1 H1 H1 H1 H1 T1 K1 T1 K+BSA-1 H+BSA1 H1 K1 T1 H1 T+BSA1 H1 H1 T1 HNot I0.5 K+BSA0.5 K+BSA1 H+BSA1 H+BSA1 H+BSA1 H+BSA0.5 K+BSA0.5 K+BSA0.5 K+BSA0.5 K+BSA1 H+BSA-1 H+BSA2 K+BSA0.5 K+BSA1 H+BSA0.5 T+BSA1 H+BSA1 H+BSA0.5 K+BSA1 H+BSAPst I1 M1 K1 H1 H1 H1 H1 M1 M1 M1 K+BSA1 H1 H+BSA-1 K1 M1 H0

6、.5 T+BSA1 H1 H1 M1 HPvu I0.5 K1 K1 K1 K1 K1 K0.5 K1 K0.5 K1 K+BSA1 K2 K+BSA1 K-0.5 K1.5 K+BSA1 K+BSA1 K1 K0.5 K1 KSac I1 M0.5 K0.5 K1 M1 M0.5 K1 M1 M1 L0.5 K+BSA1 T0.5 K+BSA1 M0.5 K-1.5 T+BSA1 T+BSA1 M0.5 K1 M1 MSal I1.5 T1.5 T1 H1 H1 H1 H1.5 K1.5 K1.5 T+BSA1.5 T+BSA1 H1 H+BSA1 H1.5 K+BSA1.5 T+BSA-1

7、.5 T+BSA1 H1 H1.5 T1 HSma I1 T+BSA0.5 T+BSA1 T+BSA1 T+BSA1 T+BSA0.5 K+BSA1 T+BSA1 T+BSA1 T+BSA1 T+BSA1 T+BSA0.5 T+BSA0.5 T+BSA1 K+BSA1 T+BSA1.5 T+BSA-1 T+BSA0.5 T+BSA1 T+BSA1 T+BSASpe I1 M1 K1 H1 M1 H1 H1 M1 M1 M1 K+BSA1 H1 H+BSA1 H1 K1 M1 H1 T+BSA-1 H1 M1 HSph I0.5 K1 K1 H1 H1 H1 H2 T1 K0.5 K1 K+BS

8、A1 H1 H+BSA1 H1 K0.5 K1 H0.5 T+BSA1 H-2 T1 HXba I1 M0.5 K2 T1 M1 M2 T1 M1 M1 M1 M+BSA1 T0.5 K+BSA1 M0.5 K1 M1.5 T1 T+BSA1 M2 T-1 MXho I1 M1 K1 H1 H1 H1 H1 M1 M1 M1 K+BSA1 H1 H+BSA1 H1 K1 M1 H1 T+BSA1 H1 H1 M-Note: 1) It is confirmed that 10 units of each enzyme completely digest 1 g of DNA at 37C in

9、 one hour in 50 l 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.Takara applies a system in whic

10、h enzyme activities measured in the optimal universal buffers (one of 5 buffers:) are displayed. The relative activity in other universal buffers is expressed as 100% of the optimal universal buffers.( ) indicates buffers which are likely to be affected by star activity etc. In order to avoid these

11、effects, use of or buffers is recommended. For 15 enzymes (Acc III, Bal I, Bcn I, Bgl I, Bpu1102 I, Cfr10 I, Eam1105 I, Eco52 I, Nru I, PshB I, SnaB I, Ssp I, Taq I or VpaK 11 BI,basal buffer specializedfor each enzyme is supplied).The compositions of basal buffers vary depending on enzymes. For ref

12、erence to double digestion, recomendable enzymes for each buffer (relative activity for each reaction mixture and enzyme) are listed.Relative activity for each reaction buffer: supplied, activity measured buffer: recommended bufferRestriction enzymeRelative activities (%)LMHKT+BSABasal*Aat II2020202

13、0100120Acc I2010020(20)16080Acc II(260)1002020200160Acc III(20)(20)20(80)(20)100Afa I60604060100100Afl II2080*2020140120Alu I1001002040200120Aor13H I20202080*80100Aor51H I801002020120120Apa I10020202020120ApaL I100202020120120Ava I(20)1002040100120Ava II801002020100100Bal I2020202040100BamH I(20)204

14、0100(20)80Ban II(120)(120)10080(100)100Bbe I2020202020100Bcn I2020406060100Bgl I2020204020100Bgl II2020100(100)(60)100Bln I20204010020120BmeT110 I20202010020140BmgT120 I20201004020240Bpu1102 I2020204060100BspT104 I100602020100120BspT107 I20208010020100Bsp1286 I10020202060100Bsp1407 I20602020100100Bs

15、sH II1001006020140100BstP I(20)(60)100(100)(100)100BstX I20401002020120Bst1107 I(20)6010010040100Cfr10 I(20)(20)(20)40(20)100Cla I4010012010060100Cpo I20208010020100Dra I100100601008080Eae I601002020120160Eam1105 I(20)(40)2040(40)100EcoO65 I(20)(60)60*4040100EcoO109 I100602020100160EcoR I(20)(100)10

16、0(120)(80)120EcoR V(20)(40)100(120)(40)100EcoT14 I(20)(40)100120(60)100EcoT22 I2020100(140)(20)120Eco52 I2020202020100Eco81 I201002020100160Fba I(20)(20)(80)100(20)100Fok I(20)(60)2020(200)100Hae II801002080140100Hae III6010010060100100Hap II10060202010080Hha I80100100120120100Hinc II20100204010080H

17、ind III(60)10020200(100)80Hinf I8010010016060100Hin1 I4080*202060160Hpa I20(40)20100(80)100Kpn I100602020(100)80Mbo I20406010040100Mbo II10060202060100Mfl I10080202080100Mlu I6060100(100)60100Msp I80802010010080Mun I(200)100*2020160100Mva I(20)(40)80100(20)120Nae I100202020100120Nco I(40)(60)2060*(6

18、0)160Nde I204010010080100Nhe I(120)1002020(160)100Not I(20)(20)20*20(20)100Nru I020202020100Nsb I40202060100100PmaC I100802020100100PshA I20402010060160PshB I(20)(40)204040100Psp1406 I20602020100100Pst I(20)(60)10080(20)80Pvu I(20)(20)(40)80*(40)120Pvu II(80)1004020(40)100Sac I1006020208080Sac II402

19、0202010040Sal I2020100(20)20120Sau 3A I(60)8010020(80)100Sca I(20)(20)100(60)(20)100Sfi I(40)1002020100100Sma I20202020100100Smi I10201004010100SnaB I(20)(40)2020(40)100Spe I(80)10080100(80)100Sph I(20)(40)100120(20)100Sse8387 I(120)60*2020(60)100Ssp I(20)(60)40(100)(80)100Stu I601006080140100Taq I4

20、080606080100Tth111 I(20)8040100(80)120Van91 I20(20)60100(60)100VpaK11B I202060(40)20100Xba I2080*2020120120Xho I206010016060100Xsp I206020100160100*+0.01% BSA100% Afl II, EcoO65 I, Fok I, Hin1 I, Mun I, Nco I, Pvu I, Spl I, Sse8387 I, Xba I*+0.01% BSA+0.01% Triton X-100100% Not I* The composition of basal buffers varies depending on enzymes.Enzymes classified regarding to universal bufferLMHKT+BSAAfa IAcc IBan IIAfa IAat IIAlu IAcc IIBgl IIAor13H I*Acc IAor51H IAfa IBmgT1