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常规内切酶反应条件


Reaction Conditions for Restriction Enzymes

Thermo Scientific Enzyme Recommended buffer for
100% activity		 Enzyme activity in 5-buffer system buffers, % Tango buffer for double digestion Units for overnight incubation,
u/μg DNA		 Thermal inactivation in 20 min
B
(blue)
1X		 G
(green)
1X		 O
(orange)
1X		 R
(red)
1X		 Tango
(yellow)
1X / 2X
AanI (PsiI) Tango 50-100 50-100 0-20 0-20 100 20-50 1X or 2X 0.5 65°C
AarI AarI +oligo NR (+oligo) NR (+oligo) 0-20 (+oligo) 0-20 (+oligo) NR (+oligo) 50-100 (+oligo) 2X (+oligo) 0.9 65°C
AatII Tango 50-100 20-50 0-20 0-20 100 20-50 1X or 2X 0.3 65°C
Acc65I (Asp718I) O 0-20 20-50 100 20-50 20-50 50-100 1X or 2X 0.3 65°C
AdeI (DraIII) G 0-20 100 20-50 100 100* 20-50 1X* or 2X 0.5 No
AjiI (BmgBI) AjiI NR NR 20-50* NR NR 20-50* 2X* 0.5 65°C
AjuI R +SAM 0-20 (+SAM) 50-100 (+SAM) 20-50 (+SAM) 100 (+SAM) 50-100 (+SAM) 50-100 (+SAM) 1X or 2X (+SAM) 0.5 65°C
AloI R 0-20 0-20 0-20 100 20-50 100 1X or 2X 0.1 65°C
AluI Tango 50-100 0-20 0-20 0-20 100 20-50 1X or 2X 0.1 65°C
Alw21I (BsiHKAI) O 0-20 20-50 100 50-100 20-50 50-100 1X or 2X 0.1 65°C
Alw26I Tango 50-100 100 0-20 0-20 100 100 1X or 2X 0.2 65°C
Alw44I Tango 50-100 100 0-20 50-100 100 50-100 1X or 2X 0.1 65°C
ApaI B 100 20-50 0-20 0-20 20-50 0-20 1X 0.2 65°C
BamHI BamHI 20-50* 100 20-50 50-100* 100* 50-100 1X* or 2X 0.5 80°C (10u)
BauI (BssSI) Tango 0-20 50-100 0-20 50-100 100 50-100 1X or 2X 0.2 65°C
BclI G 20-50 100 20-50 20-50 100* 100 1X* or 2X 0.1 80°C (10u)
BcnI (NciI) Tango 20-50 50-100 50-100 50-100 100 50-100 1X or 2X 0.2 65°C
BcuI (SpeI) Tango 50-100 50-100 0-20 20-50 100 0-20 1X 0.5 No
BfmI (SfcI) Tango 0-20 50-100 0-20 0-20 100 20-50 1X or 2X 0.9 65°C
BfuI (BciVI) BfuI NR NR 0-20 0-20 NR 50-100* 2X* 0.9 80°C
BglI O 0-20 50-100 100 100 0-20 100 2X 0.1 65°C
BglII O 0-20 20-50 100 50-100 0-20 100 2X 0.1 No
Bme1390I (ScrFI) O 20-50 50-100 100 50-100 50-100 50-100 1X or 2X 0.2 80°C
BoxI (PshAI) Tango 0-20 0-20 0-20 20-50 100 20-50 1X or 2X 0.5 80°C
BpiI (BbsI) G 20-50 100 50-100 50-100 50-100 50-100 1X or 2X 0.3 65°C
BplI Tango 0-20 (+SAM) 20-50 (+SAM) 0-20 (+SAM) 0-20 (+SAM) 100 (+SAM) 20-50 (+SAM) 1X (+SAM) 0.3 65°C
Bpu10I Bpu10I 0-20 20-50* 50-100* 100* 50-100* 100* 1X* or 2X* 0.2 80°C
Bpu1102I (BlpI) Tango 50-100 50-100 20-50 20-50 100 20-50 1X or 2X 0.1 80°C
BseDI (BsaJI) Tango 50-100 20-50 0-20 0-20 100 50-100 1X or 2X 0.2 80°C
BseGI (BtsCI) Tango 20-50 50-100 20-50 20-50 100 20-50 1X or 2X 0.2 80°C
BseJI (BsaBI) O NR 100* 100 NR NR 100* 2X* 0.1 No
BseLI (BslI) Tango 20-50 100 0-20 20-50 100 50-100 1X or 2X 0.1 No
BseMI (BsrDI) R 0-20 20-50 0-20 100 50-100 50-100 1X or 2X 0.3 80°C
BseMII (BspCNI) Tango 50-100 (+SAM) 50-100 (+SAM) 50-100 (+SAM) 50-100 (+SAM) 100 (+SAM) 50-100 (+SAM) 1X or 2X (+SAM) 0.5 80°C
BseNI (BsrI) B 100 20-50 0-20 0-20 50-100 20-50 1X or 2X 0.1 80°C
BseSI (Bme1580I) G 20-50 100 0-20 20-50 50-100 0-20 1X 0.1 80°C (10u)
BseXI (BbvI) BseXI NR NR NR NR NR NR NR 0.3 80°C
Bsh1236I (BstUI) R 0-20 0-20 50-100 100 20-50 50-100 1X or 2X 0.1 65°C
Bsh1285I (BsiEI) G 20-50 100 20-50 0-20 0-20 20-50 2X 0.2 65°C
BshNI (BanI) O 0-20 20-50 100 50-100 0-20 100 2X 0.3 65°C
BshTI (AgeI) O 0-20 20-50 100 50-100 20-50 20-50 1X or 2X 0.1 65°C
Bsp119I (BstBI) Tango 20-50 0-20 0-20 0-20 100 100 1X or 2X 0.1 80°C
Bsp120I (PspOMI) B 100 20-50 0-20 20-50 50-100 0-20 1X 0.1 80°C
Bsp1407I (BsrGI) Tango 0-20 20-50 0-20 20-50 100 50-100 1X or 2X 0.1 65°C
Bsp143I (Sau3AI) Bsp143I 20-50 20-50 0-20 0-20 50-100 20-50 1X or 2X 0.1 65°C
Bsp68I (NruI) O 0-20 20-50 100 50-100 20-50 50-100 1X or 2X 0.1 65°C
BspLI (NlaIV) Tango 50-100 50-100 0-20 20-50 100 20-50 1X or 2X 0.3 65°C
BspOI (BmtI) O 0-20 0-20 100 100 0-20 20-50 2X 0.2 80°C
BspPI (AlwI) Tango 20-50 20-50 0-20 0-20 100 0-20 1X 0.5 80°C
BspTI (AflII) O 0-20 0-20 100 20-50 0-20 50-100 2X 0.1 65°C
Bst1107I (BstZ17I) O 20-50 50-100 100 100 20-50 100 1X or 2X 0.1 65°C
BstXI O 20-50 100 100 50-100 50-100 100 1X or 2X 0.1 65°C
Bsu15I (ClaI) Tango 20-50 20-50 20-50 20-50 100 20-50 1X or 2X 0.1 65°C
BsuRI (HaeIII) R 0-20 0-20 0-20 100 50-100 100 1X or 2X 0.1 80°C
BveI (BspMI) O +oligo 0-20 (+oligo) 20-50 (+oligo) 100 (+oligo) 20-50 (+oligo) 50-100 (+oligo) 100 (+oligo) 1X or 2X (+oligo) 0.2 65°C
CaiI (AlwNI) Tango 20-50 20-50 20-50 50-100 100 50-100 1X or 2X 0.2 65°C
Cfr10I (BsrFI) Cfr10I 0-20 20-50 20-50 50-100 20-50 50-100 1X or 2X 0.1 No
Cfr13I (Sau96I) Tango 50-100 50-100 20-50 20-50 100 20-50 1X or 2X 0.3 65°C
Cfr42I (SacII) B 100 50-100 0-20 0-20 50-100 0-20 1X 0.1 65°C
Cfr9I (XmaI) Cfr9I 0-20 0-20 0-20 0-20 20-50 0-20 1X 0.2 65°C
CpoI (RsrII) Tango 20-50 50-100 50-100 20-50 100 50-100 1X or 2X 0.5 65°C
CseI (HgaI) R NR 50-100* 50-100 100 100* 50-100 1X* or 2X 0.5 80°C (10u)
Csp6I (CviQI) B 100 50-100 0-20 0-20 50-100 0-20 1X 0.1 65°C
DpnI Tango 100 100 50-100 50-100 100 50-100 1X or 2X 0.1 80°C
DraI Tango 50-100 50-100 20-50 20-50 100 50-100 1X or 2X 0.1 65°C
Eam1104I (EarI) Tango 50-100 50-100 0-20 0-20 100 0-20 1X 0.5 65°C
Eam1105I (AhdI) Eam1105I 20-50 50-100 0-20 0-20 50-100 20-50 1X or 2X 0.1 65°C
Ecl136II (EcoICRI) Ecl136II 50-100 20-50 0-20 0-20 50-100 0-20 1X 0.2 65°C
Eco105I (SnaBI) Tango 100* 50-100 0-20 0-20 100 0-20 1X 0.5 65°C
Eco130I (StyI) O 0-20 20-50 100 50-100 50-100 100 1X or 2X 0.2 65°C
Eco147I (StuI) B 100 50-100 20-50 20-50 50-100 0-20 1X 0.1 80°C
Eco24I (BanII) Tango 50-100 50-100 20-50 20-50 100 0-20 1X 0.2 65°C
Eco31I (BsaI) G 50-100 100 0-20 0-20 50-100 20-50 1X or 2X 0.3 65°C
Eco32I (EcoRV) R 0-20 50-100 50-100 100 20-50 100 1X or 2X 0.1 65°C
Eco47I (AvaII) R 0-20 50-100 50-100 100 50-100 50-100 1X or 2X 0.3 65°C
Eco47III (AfeI) O 0-20 20-50 100 100 50-100 100 1X or 2X 0.1 80°C
Eco52I (EagI) Eco52I 0-20 0-20 0-20 20-50 0-20 20-50 2X 0.2 65°C
Eco57I (AcuI) G +SAM 100 (+SAM) 100 (+SAM) 20-50 (+SAM) 20-50 (+SAM) 50-100 (+SAM) 50-100 (+SAM) 1X or 2X (+SAM) 0.9 65°C
Eco72I (PmlI) Tango NR NR 0-20 0-20 100 20-50 1X or 2X 0.5 65°C
Eco81I (Bsu36I) Tango 50-100 100 0-20 0-20 100 0-20 1X 0.1 80°C
Eco88I (AvaI) Tango 100 50-100 0-20 0-20 100 20-50 1X or 2X 0.2 65°C
Eco91I (BstEII) O 20-50 20-50 100 50-100 NR 100 2X 0.1 65°C
EcoO109I (DraII) Tango 50-100 20-50 20-50 20-50 100 100 1X or 2X 0.2 65°C
EcoRI EcoRI 0-20 NR 100 100* NR 100 2X 0.2 65°C
EcoRII O 20-50 50-100 100 50-100 20-50 50-100 1X or 2X 0.1 80°C
EheI (SfoI) Tango 20-50 50-100 0-20 0-20 100 20-50 1X or 2X 0.9 65°C
Esp3I (BsmBI) Tango 100 (+DTT) 20-50 (+DTT) 0-20 (+DTT) 0-20 (+DTT) 100 (+DDT) 0-20 (+DTT) 1X (+DDT) 0.2 65°C
FaqI (BsmFI) Tango 20-50 (+SAM) 20-50 (+SAM) 0-20 (+SAM) 0-20 (+SAM) 100 (+SAM) 50-100 (+SAM) 1X (+SAM) 0.9 80°C
FspAI O 0-20 0-20 100 50-100 0-20 50-100 2X 0.2 65°C
FspBI (BfaI) Tango 50-100 20-50 0-20 0-20 100 0-20 1X 0.3 65°C
GsuI (BpmI) B 100 50-100 0-20 20-50 100 50-100 1X or 2X 0.9 65°C
HhaI Tango 50-100 50-100 20-50 20-50 100 20-50 1X or 2X 0.1 80°C
Hin1I (BsaHI) G 20-50 100 20-50 20-50 20-50 20-50 1X or 2X 0.1 65°C
Hin1II (NlaIII) G 50-100 100 20-50 50-100 50-100 50-100 1X or 2X 0.3 65°C
Hin6I (HinP1I) Tango 50-100 50-100 20-50 20-50 100 50-100 1X or 2X 0.1 65°C
HincII (HindII) Tango 50-100 50-100 20-50 50-100 100 50-100 1X or 2X 0.1 65°C
HindIII R 0-20 20-50 0-20 100 50-100 50-100 1X or 2X 0.1 80°C
HinfI R 0-20 20-50 50-100 100 50-100 50-100 1X or 2X 0.1 65°C
HpaII Tango 50-100 50-100 0-20 20-50 100 20-50 1X or 2X 0.1 65°C
HphI B 100 0-20 0-20 0-20 20-50 0-20 1X 0.1 65°C
Hpy8I (MjaIV) Tango 50-100 50-100 0-20 20-50 100 50-100 1X or 2X 0.1 80°C
HpyF10VI (MwoI) Tango 0-20 0-20 0-20 0-20 100 50-100 1X or 2X 0.1 80°C
HpyF3I (DdeI) Tango 20-50 20-50 20-50 20-50 100 50-100 1X or 2X 0.2 65°C
I-SceI Tango 50-100 50-100 50-100 50-100 100 50-100 1X or 2X 0.9 65°C
Kpn2I (BspEI) Tango 50-100 50-100 0-20 20-50 100 50-100 1X or 2X 0.1 80°C
KpnI KpnI 20-50 0-20 0-20 0-20 20-50 0-20 1X 0.2 80°C
KspAI (HpaI) B 100 50-100* 20-50 20-50 100* 50-100 1X* or 2X* 0.5 65°C
LguI (SapI) Tango 20-50 50-100 20-50 20-50 100 20-50 1X or 2X 0.5 65°C
Lsp1109I (BbvI) Lsp1109I 0-20 20-50* 50-100* 100 20-50* 20-50* 1X* or 2X 0.5 65°C
LweI (SfaNI) Tango 0-20 0-20 0-20 20-50 100 20-50 1X or 2X 0.2 65°C
MauBI Tango 0-20 0-20 0-20 0-20 100 0-20 1X 0.1 65°C
MbiI (BsrBI) Tango 20-50 100 0-20 20-50 100 20-50 1X or 2X 0.2 65°C
MboI R 50-100 50-100 50-100 100 50-100 100 1X or 2X 0.1 65°C
MboII B 100 50-100 20-50 0-20 50-100 20-50 1X or 2X 0.9 65°C
MlsI (MscI) R 0-20 20-50 0-20 100 20-50 50-100 1X or 2X 0.5 65°C
MluI R 0-20 20-50 50-100 100 20-50 50-100 1X or 2X 0.1 65°C
MnlI G 50-100 100 20-50 20-50 20-50 20-50 1X or 2X 0.5 65°C
Mph1103I (NsiI) R 0-20 50-100 20-50 100 50-100 50-100 1X or 2X 0.3 65°C
MreI (Sse232I) G 0-20 100 0-20 0-20 50-100 0-20 1X 0.5 80°C
MspI (HpaII) Tango 50-100 50-100 0-20 0-20 100 50-100 1X or 2X 0.3 65°C
MssI (PmeI) B 100 0-20 0-20 0-20 20-50 0-20 1X 0.5 65°C
MunI (MfeI) G 100 100 0-20 0-20 100 0-20 1X 0.1 65°C
Mva1269I (BsmI) R 0-20 20-50 50-100 100 0-20 50-100 2X 0.1 65°C
MvaI (BstNI) R 20-50 20-50 50-100 100 20-50* 100 1X* or 2X 0.1 80°C (10u)
Nb.Bpu10I R 0-20 20-50 20-50 100 20-50 50-100 1X or 2X 0.3 80°C
Nb.Mva1269I O 0-20 20-50 100 100 20-50 50-100 1X or 2X 0.3 80°C
NcoI Tango 20-50 20-50 20-50 50-100 100 100 1X or 2X 0.1 65°C
NdeI O 0-20 0-20 100 50-100 0-20 50-100 2X 0.3 65°C
NheI Tango 100 20-50 0-20 0-20 100 0-20 1X 0.2 65°C
NmuCI (Tsp45I) R 0-20 20-50 50-100 100 20-50 50-100 1X or 2X 0.1 65°C
NotI O 0-20 20-50 100 20-50 0-20 20-50 2X 0.1 65°C
NsbI (FspI) Tango 20-50 50-100 0-20 20-50 100 20-50 1X or 2X 0.1 65°C
Nt.Bpu10I R 0-20 0-20 100 100 0-20 50-100 2X 0.3 80°C
OliI (AleI) R 0-20 0-20 0-20 100 0-20 50-100 2X 0.2 65°C
PacI PacI 20-50 20-50 0-20 0-20 0-20 0-20 NR 0.3 65°C
PaeI (SphI) B 100 50-100 0-20 0-20 50-100 0-20 1X 0.1 65°C
PagI (BspHI) O 0-20 50-100 100 NR NR NR NR 0.2 80°C
PasI PasI NR NR NR NR NR NR NR 0.2 80°C
PauI (BssHII) R 0-20 0-20 100 100 0-20 100 2X 0.2 80°C
PdiI (NaeI) Tango 50-100 20-50 0-20 0-20 100 50-100 1X or 2X 0.5 65°C
PdmI (XmnI) Tango 20-50 50-100 0-20 0-20 100 0-20 1X 0.5 65°C
PfeI (TfiI) O 0-20 20-50 100 50-100 20-50 50-100 1X or 2X 0.1 65°C
Pfl23II (BsiWI) Tango 20-50 50-100 20-50 20-50 100 0-20 1X 0.3 65°C
PfoI Tango 0-20 20-50 50-100 0-20 100 50-100 1X or 2X 0.1 65°C
Ppu21I (BsaAI) Ppu21I 50-100* 100* 20-50 NR NR NR NR 0.5 65°C
PscI (PciI) Tango 20-50 20-50 0-20 0-20 100 0-20 1X 0.2 65°C
Psp1406I (AclI) Tango 100 50-100 0-20 20-50 100 0-20 1X 0.5 65°C
Psp5II (PpuMI) G 0-20 100 20-50 20-50 50-100 100 1X or 2X 0.2 80°C
PstI O 50-100 50-100 100 100 50-100 50-100 1X or 2X 0.2 80°C (10u)
PsuI (BstYI) B 100 20-50 0-20 0-20 50-100 0-20 1X 0.5 80°C
PsyI (Tth111I) B 100 50-100 0-20 0-20 50-100 0-20 1X 0.1 80°C
PvuI R 0-20 20-50 50-100 100 50-100 100 1X or 2X 0.2 80°C (10u)
PvuII G 50-100* 100 20-50 50-100 20-50* 20-50* 1X* or 2X* 0.2 80°C
RsaI Tango 50-100 20-50 0-20 0-20 100 0-20 1X 0.2 65°C
RseI (MslI) R 0-20 50-100 50-100 100 20-50 100 1X or 2X 0.5 65°C
SacI SacI 50-100 20-50 0-20 0-20 50-100 20-50 1X or 2X 0.2 65°C
SalI O 0-20 0-20 100 20-50 0-20 50-100 2X 0.1 65°C
SatI (Fnu4HI) G 20-50 100 20-50 20-50 50-100 20-50 1X or 2X 0.1 65°C
ScaI ScaI 0-20 0-20 0-20 0-20 0-20 0-20 NR 0.3 80°C (10u)
SchI (MlyI) Tango 20-50 50-100 0-20 0-20 100 0-20 1X 0.2 65°C
SdaI (SbfI) SdaI NR NR 0-20 0-20 NR 20-50 2X 0.3 65°C
SduI (Bsp1286I) SduI NR 50-100* 0-20 0-20 NR NR NR 0.3 65°C
SfaAI (AsiSI) Tango 50-100 0-20 0-20 0-20 100 0-20 1X 0.2 80°C
SfiI G 50-100 100 20-50 0-20 100 0-20 1X 0.2 80°C
SgeI SgeI 0-20 0-20 0-20 NR NR NR NR 0.0 65°C
SgrDI R 0-20 0-20 0-20 100 NR 100 2X 0.3 65°C
SgsI (AscI) Tango 0-20 0-20 0-20 50-100 100 50-100 1X or 2X 0.1 65°C
SmaI Tango 50-100 0-20 0-20 0-20 100 0-20 1X 0.2 65°C
SmiI (SwaI) O 0-20 0-20 100 20-50 0-20 20-50 2X 0.1 65°C
SmoI (SmlI) Tango 50-100 20-50 0-20 20-50 100 20-50 1X or 2X 0.2 80°C
SsiI (AciI) O NR 20-50 100 50-100 NR 100 2X 0.5 65°C
SspDI (KasI) Tango 20-50 0-20 NR 0-20 100 20-50 1X or 2X 1.0 80°C
SspI G 20-50 100 0-20 50-100 100 20-50 1X or 2X 0.1 65°C
TaaI (HpyCH4III) Tango 0-20 0-20 0-20 50-100 100 100 1X or 2X 0.2 No
TaiI (MaeII) R 50-100 50-100 20-50 100 100 50-100 1X or 2X 0.3 No
TaqI TaqI 0-20 20-50 20-50 20-50 20-50 20-50 1X or 2X 0.3 No
TasI (Tsp509I) B 100 50-100 20-50 0-20 20-50 0-20 1X 0.3 No
TatI Tango NR 50-100* 0-20 20-50 100* 0-20 1X* 0.2 No
TauI B 100 50-100 0-20 0-20 20-50 0-20 1X 0.9 No
Tru1I (MseI) R 50-100 50-100 20-50 100 50-100 100 1X or 2X 0.2 No
TscAI (TspRI) Tango 50-100 50-100 20-50 20-50 100 20-50 1X or 2X 0.2 No
Van91I (PflMI) R 0-20 50-100 50-100 100 20-50 50-100 1X or 2X 0.1 65°C
VspI (AseI) O 0-20 50-100 100 20-50 100 100 1X or 2X 0.1 65°C
XagI (EcoNI) R 0-20 20-50 50-100 100 20-50 50-100 1X or 2X 0.1 65°C
XapI (ApoI) Tango 50-100 100 0-20 0-20 100 20-50 1X or 2X 0.1 65°C
XbaI Tango 50-100 50-100 20-50 0-20 100 50-100 1X or 2X 0.1 65°C
XceI (NspI) Tango 50-100 0-20 0-20 0-20 100 0-20 1X 0.2 65°C
XhoI R 0-20 50-100 50-100 100 20-50 100 1X or 2X 0.1 80°C
XmaJI (AvrII) Tango 20-50 50-100 50-100 50-100 100 50-100 1X or 2X 0.2 80°C
XmiI (AccI) B 100 0-20 0-20 0-20 50-100 20-50 1X or 2X 0.1 65°C