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