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NEBuffer Activity Chart for Restriction Enzymes

For your convenience, New England Biolabs offers a simple 4 buffer system. A color-coded 10X NEBuffer is supplied with every restriction endonuclease, ensuring 100% activity. On November 1, 2008, NEB changed the recommended buffer for 54 restriction enzymes to NEBuffer 4. With this change, NEB now provides over 160 restriction enzymes in a single buffer. This results in increased efficiency, flexibility and ease of use especially when performing double digests.

To help select the best conditions for double digests, this chart shows the optimal (supplied) NEBuffer and approximate activity in the four standard NEBuffers and NEBuffer EcoRI for each enzyme. If BSA is supplied with an enzyme, it is included in all NEBuffer activity reactions. In addition, the table also shows recommended reaction temperature, heat inactivation temperature, recommended diluent buffer and whether the enzyme is Time-Saver qualified (i.e., cleaves substrate in 5 minutes).

Click for the Chart Legend, Icon Description

A | B | C | D | E | F | H | I | K | M | N | P | R | S | T | X | Z |

Enzyme	Supplied NEBuffer	% Activity in NEBuffers					Heat Inac. °C	Incu. Temp. °C	Dilu- ent
Enzyme	Supplied NEBuffer	1	2	3	4	EcoRI	Heat Inac. °C	Incu. Temp. °C	Dilu- ent
AatII	NEBuffer 4	0	50	50	100	25	65	37	A
Acc65I	NEBuffer 3 + BSA	10	75	100	25	50	65	37	A
AccI	NEBuffer 4	50	50	10	100	5	80	37	A
AciI	NEBuffer 3	25	50	100	50	100	65	37	A
AclI	NEBuffer 4 + BSA	10	10	0	100	5	No	37	B
AcuI	NEBuffer 4 + SAM	50	100	50	100	100	65	37	B
AfeI	NEBuffer 4	25	50	25	100	100	65	37	B
AflII	NEBuffer 4 + BSA	50	100	25	100	0	65	37	A
AflIII	NEBuffer 3 + BSA	25	75	100	50	100	80	37	B
AgeI	NEBuffer 1	100	50	10	75	50	65	37	C
AgeI-HF&trade	NEBuffer 4 + BSA	100	50	10	100	-	65	37	A
AhdI	NEBuffer 4 + BSA	25	75	0	100	10	65	37	A
AleI	NEBuffer 4	10	25	10	100	0	65	37	A
AluI	NEBuffer 4	100	100	75	100	100	65	37	B
AlwI	NEBuffer 4	50	100	10	100	5	65	37	A
AlwNI	NEBuffer 4	50	100	50	100	10	65	37	A
ApaI	NEBuffer 4 + BSA	25	50	0	100	5	65	25	A
ApaLI	NEBuffer 4 + BSA	100	100	10	100	5	No	37	A
ApeKI	NEBuffer 3	25	75	100	50	100	No	75	B
ApoI	NEBuffer 3 + BSA	10	75	100	75	100	80	50	A
AscI	NEBuffer 4	0	10	10	100	5	65	37	A
AseI	NEBuffer 3	NR	75	100 ^dd	NR	50	65	37	B
AsiSI	NEBuffer 4 + BSA	50	100	100	100	0	80	37	B
AvaI	NEBuffer 4	10	75	10	100	50	80	37	A
AvaII	NEBuffer 4	50	75	10	100	50	65	37	A
AvrII	NEBuffer 4	100	100	50	100	50	80	37	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
BaeGI	NEBuffer 1	100	75	10	50	100	65	37	A
BaeI	NEBuffer 4 + BSA + SAM	50	100	50	100	40	65	25	A
BamHI	NEBuffer 3 + BSA	75	100	100 ^dd	100	100	No	37	A
BamHI-HF^&trade	NEBuffer 4	100	50	10	100	25	No	37	A
BanI	NEBuffer 4 + BSA	50	100	50	100	25	65	37	A
BanII	NEBuffer 4	100	10	50	100	100	80	37	A
BbsI	NEBuffer 2	100	100	25	75	50	65	37	B
BbvCI	NEBuffer 4	50	100	10	100	100	80	37	A
BbvI	NEBuffer 2	100	100	25	75	100	65	37	B
BccI	NEBuffer 1 + BSA	100	50	10	50	15	65	37	A
BceAI	NEBuffer 3 + BSA	100	100	100	100	100	65	37	A
BcgI	NEBuffer 3 + SAM	NR	NR	100 ^dd	NR	50	65	37	A
BciVI	NEBuffer 4	100	50	0	100	25	65	37	C
BclI	NEBuffer 3	50	100	100 ^dd	75	100	No	50	A
BfaI	NEBuffer 4	75	50	10	100	0	80	37	B
BfuAI	NEBuffer 3	0	75	100	10	100	65	50	B
BfuCI	NEBuffer 4 + BSA	100	50	25	100	15	80	37	B
BglI	NEBuffer 3	50	75	100	50	100	65	37	B
BglII	NEBuffer 3	10	75	100	10	100	No	37	A
BlpI	NEBuffer 4	50	100	10	100	50	No	37	A
BmgBI	NEBuffer 3 + BSA	0	25	100	10	50	65	37	C
BmrI	NEBuffer 2	75	100	75	100	50	65	37	B
BmtI	NEBuffer 2	25	100	25	50	100	65	37	B
BpmI	NEBuffer 3 + BSA	75	100	100	100	100	65	37	B
Bpu10I	NEBuffer 3	10	25	100	25	100	80	37	B
BpuEI	NEBuffer 4 + SAM	50	100	10	100	100	65	37	B
BsaAI	NEBuffer 4	100	100	100	100	100	No	37	C
BsaBI	NEBuffer 4	50	100	75	100	100	80	60	B
BsaHI	NEBuffer 4 + BSA	50	100	100	100	100	80	37	A
BsaI	NEBuffer 4 + BSA	75	75	100	100	100	65	37	B
BsaI-HF&trade	NEBuffer 4 + BSA	25	25	10	100	-	65	37	B
BsaJI	NEBuffer 4	100	100	100	100	100	80	60	A
BsaWI	NEBuffer 4 + BSA	50	100	50	100	40	80	60	A
BsaXI	NEBuffer 4	75	100	10	100	15	No	37	B
BseRI	NEBuffer 4	100	100	75	100	100	65	37	A
BseYI	NEBuffer 3	10	50	100	50	100	65	37	B
BsgI	NEBuffer 4 + SAM	50	75	50	100	20	65	37	B
BsiEI	NEBuffer 4 + BSA	50	100	10	100	5	80	60	A
BsiHKAI	NEBuffer 4 + BSA	50	100	100	100	25	No	65	B
BsiWI	NEBuffer 3	100	100	100	25	100	80	55	A
BslI	NEBuffer 3	10	50	100	75	-	80	55	A
BsmAI	NEBuffer 4	50	100	100	100	100	80	55	B
BsmBI	NEBuffer 3	75	100	100	100	100	80	55	B
BsmFI	NEBuffer 4 + BSA	25	50	50	100	15	80	65	A
BsmI	NEBuffer 4	75	100	75	100	5	80	65	A
BsoBI	NEBuffer 2	10	100	100	50	100	No	37	A
Bsp1286I	NEBuffer 4 + BSA	25	25	25	100	10	65	37	A
BspCNI	NEBuffer 4 + BSA + SAM	100	75	10	100	15	80	25	A
BspDI	NEBuffer 4	25	75	50	100	100	65	37	A
BspEI	NEBuffer 3	0	10	100	0	100	80	37	B
BspHI	NEBuffer 4	10	100	50	100	5	65	37	A
BspMI	NEBuffer 3	NR	NR	100	NR	100	65	37	B
BspQI	NEBuffer 4	10	50	100	100	100	80	50	B
BsrBI	NEBuffer 4	50	100	100	100	100	80	37	A
BsrDI	NEBuffer 2 + BSA	50	100	50	75	50	80	65	A
BsrFI	NEBuffer 4	10	100	100	100	100	No	37	C
BsrGI	NEBuffer 2 + BSA	25	100	10	100	50	80	37	A
BsrI	NEBuffer 3	0	50	100	10	100	80	65	B
BssHII	NEBuffer 3	100	100	100 ^dd	100	100	80	50	A
BssKI	NEBuffer 3 + BSA	0	50	100	50	100	80	60	A
BssSI	NEBuffer 3 + BSA	0	50	100 ^dd	10	100	80	37	B
BstAPI	NEBuffer 4 + BSA	25	100	100	100	100	80	60	A
BstBI	NEBuffer 4	75	50	25	100	50	No	65	A
BstEII	NEBuffer 3 + BSA	50	75	100	75	100	No	60	A
BstNI	NEBuffer 2 + BSA	10	100	100	75	50	No	60	A
BstUI	NEBuffer 4	100	100	50	100	100	No	60	A
BstXI	NEBuffer 3	0	50	100	25	100	65	37	B
BstYI	NEBuffer 2	50	100	75	100	50	80	60	A
BstZ17I	NEBuffer 4	NR	NR	100	100	100	No	37	B
Bsu36I	NEBuffer 3 + BSA	0	25	100	0	15	80	37	A
BtgI	NEBuffer 3 + BSA	25	50	100	100	100	80	37	B
BtgZI	NEBuffer 4 + BSA	10	25	0	100	50	80	60	A
BtsCI	NEBuffer 4	50	100	50	100	25	No	50	B
BtsI	NEBuffer 4 + BSA	100	50	50	100	25	80	55	A
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
Cac8I	NEBuffer 4	50	75	100	100	50	65	37	B
ClaI	NEBuffer 4 + BSA	10	50	50	100	50	65	37	A
CspCI	NEBuffer 4 + SAM	10	100	10	100	20	65	37	A
CviAII	NEBuffer 4	75	25	10	100	25	65	25	B
CviKI-1	NEBuffer 4	10	50	25	100	100	80	37	A
CviQI	NEBuffer 3 + BSA	75	100	100	75	0	No	25	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
DdeI	NEBuffer 3	75	100	100	75	100	65	37	A
DpnI	NEBuffer 4	100	100	75	100	100	80	37	B
DpnII	NEBuffer DpnII	NR	NR	100 ^dd	NR	100	65	37	B
DraI	NEBuffer 4	75	75	50	100	100	65	37	A
DraIII	NEBuffer 3 + BSA	100	75	100	25	50	65	37	B
DrdI	NEBuffer 4 + BSA	25	50	10	100	100	65	37	A
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
EaeI	NEBuffer 4	100	100	50	100	100	80	37	A
EagI	NEBuffer 3	10	25	100	10	100	65	37	C
EagI-HF^&trade	NEBuffer 4	25	50	10	100	100	65	37	B
EarI	NEBuffer 4	100	100	50	100	100	65	37	A
EciI	NEBuffer 2 + BSA	50	100	100	50	100	65	37	A
Eco53kI	NEBuffer 4	100	50	25	100	-	65	37	A
EcoNI	NEBuffer 4	100	100	75	100	100	65	37	A
EcoO109I	NEBuffer 4 + BSA	100	100	75	100	15	65	37	A
EcoP15I	NEBuffer 3 + BSA + ATP	75	100	100	100	50	65	37	A
EcoRI	NEBuffer EcoRI ^dd	100	100	100	100	100	65	37	C
EcoRI-HF^&trade	NEBuffer 4	10	100	0	100	5	65	37	C
EcoRV	NEBuffer 3 + BSA	50	75	100	50	50	80	37	A
EcoRV-HF^&trade	NEBuffer 4	25	100	100	100	100	65	37	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
FatI	NEBuffer 2	10	100	50	50	100	65	55	A
FauI	NEBuffer 4	100	50	10	100	50	65	55	A
Fnu4HI	NEBuffer 4	10	25	25	100	10	65	37	A
FokI	NEBuffer 4	100	100	75	100	25	65	37	A
FseI	NEBuffer 4 + BSA	100	75	0	100	50	65	37	B
FspI	NEBuffer 4	10	75	10	100 ^dd	50	65	37	C
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
HaeII	NEBuffer 4 + BSA	75	100	50	100	25	80	37	A
HaeIII	NEBuffer 4	50	100	25	100	100	80	37	A
HgaI	NEBuffer 1	100	75	50	100	25	65	37	A
HhaI	NEBuffer 4 + BSA	75	100	100	100	100	65	37	A
HincII	NEBuffer 3 + BSA	75	100	100	100	100	65	37	B
HindIII	NEBuffer 2	50	100	10	50	25	65	37	B
HinfI	NEBuffer 4	75	100	75	100	100	80	37	A
HinP1I	NEBuffer 4	100	100	100	100	100	65	37	A
HpaI	NEBuffer 4	25	50	10	100	100	No	37	A
HpaII	NEBuffer 1	100	50	10	50	15	65	37	A
HphI	NEBuffer 4	NR	75	0	100	10	65	37	B
Hpy166II	NEBuffer 4	100	100	50	100	20	65	37	C
Hpy188I	NEBuffer 4	50	25	10	100	50	65	37	A
Hpy188III	NEBuffer 4 + BSA	100	100	10	100	5	65	37	A
Hpy99I	NEBuffer 4 + BSA	10	10	0	100	25	65	37	A
HpyAV	NEBuffer 4 + BSA	100	100	25	100	50	65	37	B
HpyCH4III	NEBuffer 4	100	50	25	100	20	80	37	A
HpyCH4IV	NEBuffer 1	100	25	10	25	15	65	37	A
HpyCH4V	NEBuffer 4	50	50	25	100	50	65	37	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
I-CeuI	NEBuffer 4 + BSA	10	10	0	100	-	65	37	B
I-SceI	NEBuffer I-SceI + BSA	10	50	50	50	-	65	37	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
KasI	NEBuffer 4 + BSA	25	100	0	100	100	65	37	B
KpnI	NEBuffer 1 + BSA	100	75	0	50	25	No	37	A
KpnI-HF&trade	NEBuffer 4	100	25	0	100	-	No	37	A
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
MboI	NEBuffer 4	75	100	100	100	100	65	37	A
MboII	NEBuffer 4	100	100	50	100	100	65	37	C
MfeI	NEBuffer 4	75	50	10	100	5	65	37	A
MfeI-HF^&trade	NEBuffer 4	75	50	10	100	0	65	37	A
MluI	NEBuffer 3	25	75	100	50	100	65	37	A
MlyI	NEBuffer 4 + BSA	50	50	25	100	25	65	37	A
MmeI	NEBuffer 4 + SAM	NR	NR	NR	100 ^dd	50	80	37	B
MnlI	NEBuffer 4 + BSA	75	100	50	100	50	65	37	B
MscI	NEBuffer 4	75	75	75	100	100	65	37	B
MseI	NEBuffer 4 + BSA	75	100	75	100	50	65	37	A
MslI	NEBuffer 4	100	100	25	100	10	65	37	A
MspA1I	NEBuffer 4 + BSA	50	100	50	100	50	65	37	B
MspI	NEBuffer 4	75	100	50	100	50	80	37	A
MwoI	NEBuffer 3	10	75	100	75	100	No	60	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
NaeI	NEBuffer 4	100	75	10	100	25	65	37	A
NarI	NEBuffer 4	100	75	75	100	50	65	37	A
Nb.BbvCI	NEBuffer 2	50	100	10	100	-	80	37	A
Nb.BbvCI	NEBuffer 2	50	100	10	100	-	80	37	A
Nb.BsmI	NEBuffer 3	25	100	100	25	-	80	65	A
Nb.BsmI	NEBuffer 3	25	100	100	25	-	80	65	A
Nb.BsrDI	NEBuffer 2	10	100	25	50	-	80	65	A
Nb.BsrDI	NEBuffer 2	10	100	25	50	-	80	65	A
Nb.BtsI	NEBuffer 4 + BSA	75	100	75	100	-	80	37	A
Nb.BtsI	NEBuffer 4 + BSA	75	100	75	100	-	80	37	A
NciI	NEBuffer 4	100	25	10	100	25	No	37	A
NcoI	NEBuffer 3	100	100	100	100	100	65	37	A
NcoI-HF^&trade	NEBuffer 4	50	100	10	100	100	80	37	B
NdeI	NEBuffer 4	75	100	75	100	100	65	37	A
NgoMIV	NEBuffer 4	100	50	10	100	0	80	37	A
NheI	NEBuffer 2 + BSA	100	100	10	100	15	65	37	C
NheI-HF^&trade	NEBuffer 4 + BSA	100	10	0	100	0	80	37	C
NlaIII	NEBuffer 4 + BSA	25	25	25	100	5	65	37	B
NlaIV	NEBuffer 4 + BSA	10	10	10	100	100	65	37	B
NmeAIII	NEBuffer 4 + SAM	10	10	0	100	0	80	37	B
NotI	NEBuffer 3 + BSA	0	50	100	25	100	65	37	C
NotI-HF^&trade	NEBuffer 4 + BSA	25	100	25	100	50	65	37	A
NruI	NEBuffer 3	0	10	100	10	100	65	37	A
NsiI	NEBuffer 3	10	75	100	25	100	80	37	A
NspI	NEBuffer 2 + BSA	100	100	0	100	5	65	37	A
Nt.AlwI	NEBuffer 2	25	100	25	50	-	80	37	A
Nt.AlwI	NEBuffer 2	25	100	25	50	-	80	37	A
Nt.BbvCI	NEBuffer 4	50	100	10	100	-	80	37	A
Nt.BbvCI	NEBuffer 4	50	100	10	100	-	80	37	A
Nt.BsmAI	NEBuffer 4	100	50	10	100	-	65	37	A
Nt.BsmAI	NEBuffer 4	100	50	10	100	-	65	37	A
Nt.BspQI	NEBuffer 3	10	75	100	25	-	80	50	B
Nt.BspQI	NEBuffer 3	10	75	100	25	-	80	50	B
Nt.BstNBI	NEBuffer 3	0	10	100	10	-	80	55	A
Nt.BstNBI	NEBuffer 3	0	10	100	10	-	80	55	A
Nt.CviPII	NEBuffer 4	25	100	50	100	-	65	37	A
Nt.CviPII	NEBuffer 4	25	100	50	100	-	65	37	A
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
PacI	NEBuffer 1 + BSA	100	75	10	100	5	65	37	A
PaeR7I	NEBuffer 4	25	100	10	100	50	No	37	A
PciI	NEBuffer 3	50	75	100	50	0	80	37	A
PflFI	NEBuffer 4 + BSA	0	25	25	100	-	65	37	A
PflMI	NEBuffer 3 + BSA	0	100	100	50	100	65	37	A
PhoI	NEBuffer 3	50	50	100	75	100	No	75	A
PI-PspI	NEBuffer PI-PspI + BSA	0	10	10	10	-	No	65	B
PI-SceI	NEBuffer PI-SceI + BSA	0	0	0	0	-	65	37	B
PleI	NEBuffer 4	100	100	75	100	50	65	37	A
PmeI	NEBuffer 4 + BSA	0	50	10	100	5	65	37	A
PmlI	NEBuffer 1 + BSA	100	75	0	75	15	65	37	A
PpuMI	NEBuffer 4	0	25	0	100	10	No	37	B
PshAI	NEBuffer 4 + BSA	50	75	10	100	5	65	37	A
PsiI	NEBuffer 4	10	100	10	100	10	65	37	B
PspGI	NEBuffer 4	75	100	50	100	100	No	75	A
PspOMI	NEBuffer 4	25	25	10	100	15	65	37	B
PspXI	NEBuffer 4	0	100	10	100	50	80	37	B
PstI	NEBuffer 3 + BSA	75	75	100	50	50	80	37	C
PstI-HF&trade	NEBuffer 4	10	75	50	100	-	80	37	C
PvuI	NEBuffer 3 + BSA	10	75	100	10	100	80	37	B
PvuII	NEBuffer 2	100	100	100	100	100	No	37	B
PvuII-HF^&trade	NEBuffer 4	0	25	0	100	0	80	37	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
RsaI	NEBuffer 4	100	100	50	100	15	65	37	A
RsrII	NEBuffer 4	25	75	10	100	25	65	37	C
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
SacI	NEBuffer 1 + BSA	100	50	10	100	15	65	37	A
SacI-HF^&trade	NEBuffer 4 + BSA	100	50	10	100	15	65	37	A
SacII	NEBuffer 4	25	75	10	100	100	65	37	A
SalI	NEBuffer 3 + BSA	0	0	100	0	50	65	37	A
SalI-HF^&trade	NEBuffer 4	10	100	100	100	100	65	37	A
SapI	NEBuffer 4	75	50	0	100	50	65	37	C
Sau3AI	NEBuffer 1 + BSA	100	50	10	100	5	65	37	A
Sau96I	NEBuffer 4	50	100	100	100	50	80	37	A
SbfI	NEBuffer 4	75	50	0	100	25	65	37	A
SbfI-HF^&trade	NEBuffer 4	50	25	0	100	0	65	37	B
ScaI	NEBuffer 3	NR	NR	100 ^dd	NR	50	80	37	A
ScaI-HF^&trade	NEBuffer 4	100	100	10	100	15	65	37	B
ScrFI	NEBuffer 4	100	100	100	100	100	65	37	C
SexAI	NEBuffer 4 + BSA	100	75	50	100	50	65	37	A
SfaNI	NEBuffer 3	0	75	100	25	50	65	37	B
SfcI	NEBuffer 4 + BSA	75	50	10	100	25	65	37	B
SfiI	NEBuffer 4 + BSA	0	100	10	100	15	No	50	C
SfoI	NEBuffer 4	25	100	50	100	100	No	37	B
SgrAI	NEBuffer 4	100	50	10	100	50	65	37	A
SmaI	NEBuffer 4	0	0	0	100	0	65	25	A
SmlI	NEBuffer 4 + BSA	25	75	25	100	50	No	55	A
SnaBI	NEBuffer 4 + BSA	25	50	25	100 ^dd	5	80	37	A
SpeI	NEBuffer 4 + BSA	75	100	25	100	50	80	37	C
SphI	NEBuffer 2	100	100	50	100	50	65	37	B
SphI-HF^&trade	NEBuffer 4	50	25	10	100	0	65	37	B
SspI	NEBuffer SspI	50	100	50	50	100	65	37	C
SspI-HF^&trade	NEBuffer 4	25	100	0	100	25	65	37	B
StuI	NEBuffer 4	100	100	50	100	50	65	37	A
StyD4I	NEBuffer 2	10	100	100	100	20	65	37	B
StyI	NEBuffer 3 + BSA	25	75	100	10	15	65	37	A
StyI-HF&trade	NEBuffer 4	25	100	25	100	-	65	37	A
SwaI	NEBuffer 3 + BSA	10	10	100	10	15	65	25	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
Taq^αI	NEBuffer 4 + BSA	50	75	100	100	-	80	65	B
TfiI	NEBuffer 4	100	100	100	100	100	No	65	C
TliI	NEBuffer 3 + BSA	25	25	100	10	50	No	75	B
TseI	NEBuffer 4	75	100	100	100	100	No	65	B
Tsp45I	NEBuffer 1 + BSA	100	25	0	75	50	No	65	A
Tsp509I	NEBuffer 1	100	100	100	NR	100	No	65	A
TspMI	NEBuffer 4	50	75	50	100	20	No	75	B
TspRI	NEBuffer 4 + BSA	25	50	25	100	5	No	65	B
Tth111I	NEBuffer 4	50	25	25	100	100	No	65	B
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
XbaI	NEBuffer 4 + BSA	0	100	75	100	15	65	37	A
XcmI	NEBuffer 2	10	100	50	50	20	65	37	C
XhoI	NEBuffer 4 + BSA	75	100	100	100	100	65	37	A
XmaI	NEBuffer 4 + BSA	25	50	0	100	25	65	37	A
XmnI	NEBuffer 4 + BSA	100	100	50	100	5	65	37	A
Enzyme top	Supplied NEBuffer	B1	B2	B3	B4	EcoRI	Heat Inac.	Incu. Temp.	Dilu- ent
ZraI	NEBuffer 4	100	25	10	100	10	No	37	B

Note:
1. SspI Unique Buffer has the same composition as EcoRI Unique Buffer.
2. The values listed in this table are approximate. They were obtained using each enzyme's specific unit assay substrate DNA.

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