Takara_双酶切buffer用表

1、EnzymeAccIBamHIBglIIClaIEcoRIEcoRVHincIIHindIIIKpnSuppliedBuffer10M10>K10汨10M10汨10>H10M10M10AccI一0.5XK1河1>M1M0.5XK1>M1M1BamHI0.5XK一1>K1*1>K1>K0.5XK1>K0.51BglII1XT1XK一1XH1XH1XH2XK1XKClaI1XM1XK1XH一1XH1XH1Ml1XM1EcoRI1XM1XK1XH1XH一1XH1Ml1XM1EcoRV0.5XK1XK1XH1XH1XH一2XT1XK0.5HincII1X

2、M0.5XK2XK1XM1XM2XT一1XM1HindIII1XM1XK1XK1XM1XM1XK1Ml一1KpnI1XM0.5XK1XT1XM1XM0.5XK1Ml1XM-NcoI1XM+BSA1XK+BSA1XK+BSA1XK+BSA1XK+BSA1XK+BSA1XM+BSA1XK+BSA0.5+B10.5+BNdeI1XT1XK1XH1XH1XH1XH1XT1XKNotI0.5XK+BSA0.5XK+BSA1XH+BSA1XH+BSA1XH+BSA1XH+BSA0.5XK+BSA0.5>K+BSAPstI1XM1XK1XH1XH1XH1XH1Ml1XM1PvuI0.5XK1XK1XK

3、1XK1XK1XK0.5XK1XK0.5SacI1XM0.5XK0.5XK1XM1XM0.5XK1Ml1XM1SalI1.5对1.5对1XH1XH1XH1XH1_1.5XK1.5>K1.5+BSmaI1XT+BSA0.5>T+BSA1XT+BSA1XT+BSA1XT+BSA0.5XK+BSA1XT+BSA1XT+BSA1+BSpeI1XM1XK1XH1XM1XH1XH1Ml1XM1SphI0.5XK1XK1XH1XH1XH1XH2XT1XK0.5XbaI1XM0.5XK2XT1XM1XM2XT1H1XM1XhoI1XM1XK1XH1XH1XH1XH1H1XM1Note:1) Iti

4、sconfirmedthat10unitsofeachenzymecompletelydigest1gofDNAjat37Cinonehourin50lreactionmixture.2) TheconcentrationofGlycerolshouldbelessthan10%tominimizestaractivity.3) DNAmaynotbedigestedcompletely,whenrecognitionsequencesoftwoenzymesarecloseeachother,orwhenDNAtakeshigh-structureconformation.Relativea

5、ctivityforeachreactionbuffer:supplied,activitymeasuredbuffer:recommendedbufferRestrictionenzymeRelativeactivities(%)LMHKT+BSABasal*AatII<20<20<20<20100120AccI20100<20(<20)16080AccII(260)100<2020200160AccIII(<20)(<20)20(80)(<20)100AfaI60604060100100AflII2080*<20<20

6、140120AluI100100<2040200120Aor13HI<2020<20*8080100Aor51HI80100<2020120120ApaI100<20<20<20<20120ApaLI10020<20<20120120AvaI(<20)1002040100120AvaII80100<2020100100BalI2020<20<2040100BamHI(<20)<2040100(<20)80BanII(120)(120)10080(100)100BbeI<20<20&

7、lt;20<20<20100BcnI<2020406060100BglI<20<202040<20100BglII<2020100(100)(60)100BlnI<20204010020120BmeT110I<20<2020100<20140BmgT120I<20<2010040<20240Bpu1102I<20<20<204060100BspT104I10060<20<20100120BspT107I<20208010020100Bsp1286I10020<20<

8、;2060100Bsp1407I20602020100100BssHII1001006020140100BstPI(<20)(60)100(100)(100)100BstXI<2040100<20<20120Bst1107I(<20)6010010040100Cfr10I(<20)(<20)(<20)40(20)100ClaI4010012010060100CpoI<20<2080100<20100DraI100100601008080EaeI60100<20<20120160Eam1105I(<20)(40)

9、2040(40)100EcoO65I(20)(60)*604040100EcoO109I10060<20<20100160EcoRI(20)(100)100(120)(80)120EcoRV(<20)(40)100(120)(40)100EcoT14I(<20)(40)100120(60)100EcoT22I<2020100(140)(20)120Eco52I<20<20<20<20<20100Eco81I<20100<20<20100160FbaI(<20)(<20)(80)100(20)100FokI

10、(20)(60)<20<20(200)100HaeII80100<2080140100HaeIII6010010060100100HapII10060<20<2010080HhaI80100100120120100HincII20100204010080HindIII(60)100<20200(100)80HinfI8010010016060100Hin1I4080*<202060160HpaI<20(40)20100(80)100KpnI10060<20<20(100)80MboI20406010040100MboII10060&l

11、t;20<2060100MflI10080<20<2080100MluI6060100(100)60100MspIMunI80(200)80100*<20<20100<2010016080100MvaI(<20)(40)80100(20)120NaeI100<20<20<20100120NcoI(40)(60)2060*(60)160NdeI<204010010080100NheI(120)100<20<20(160)100NotI(<20)(<20)20*<20(<20)100NruI0&l

12、t;202020<20100NsbI4020<2060100100PmaCI10080<20<20100100PshAI2040<2010060160PshBI(20)(40)204040100Psp1406I2060<20<20100100PstI(<20)(60)10080(20)80PvuI(<20)(20)(40)80*(40)120PvuII(80)10040<20(40)100SacI10060<20<208080SacII4020<20<2010040SalI<20<20100(20)

13、<20120Sau3AI(60)80100<20(80)100ScaI(<20)(<20)100(60)(<20)100SfiI(40)100<20<20100100SmaI<20<20<20<20100100SmiI<10<2010040<10100SnaBI(20)(40)<20<20(40)100SpeI(80)10080100(80)100SphI(20)(40)100120(20)100Sse8387I(120)60*<20<20(60)100SspI(<20)(60)4

14、0(100)(80)100StuI601006080140100TaqI4080606080100Tth111I(20)8040100(80)120Van91I<20(20)60100(60)100VpaK11BI<20<2060(40)<20100XbaI<2080*20<20120120XhoI<206010016060100XspI<2060<20100160100Compositionsofuniversalbuffer10XL100mMTris-HCl(pH7.5)100mMMgCl210mMDithiothreitol2.10X

15、M100mMTris-HCl(pH7.5)100mMMgCl210mMDithiothreitol500mMNaCl3.10XH500mMTris-HCl(pH7.5)100mMMgCl210mMDithiothreitol1,000mMNaCl4.10XK200mMTris-HCl(pH8.5)100mMMgCl210mMDithiothreitol1,000mMKCl5.10XT(BSA-free)330mMTris-acetate(pH7.9)100mMMg-acetate5mMDithiothreitol660mMK-acetate6.0.1%BSAsolvedinsterilized

16、water7.0.1%TritonX-100InstructionsforuseUniversalBuffersare10concentration.Pleaseadd1/10volumeofthereactionmixture.Since10kbufferdoesnotcontainBSA,besuretoaddBSAtoafinalconcentrationof0.01%.Somerestrictionenzymesaresupposedtoexhibita100%activitywhenBSAorTritonX-100isaddedtothereactionsystem.SinceBSAorTritonX-100suppliedwithenzymesis10concentration(0.1%),besuretoadd1/10volumeofthereaction