NEBuffer Activity Chart for Restriction Enzymes, New England Biolabs
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Home > Technical Reference > Restriction Endonucleases > NEBuffer Activity Chart for Restriction Enzymes
NEBuffer Activity Chart for Restriction Enzymes
New England Biolabs provides a color-coded 10X NEBuffer with every restriction endonuclease to ensure 100% activity. To help select the best
conditions for double digests, this chart shows the optimal (supplied) NEBuffer and approximate activity in the four standard NEBuffers for each
enzyme. Incubation temperature is 37°C unless otherwise noted. If BSA is supplied with an enzyme, it is included in all NEBuffer activity
reactions. Additional double digestion information can be found here.
Note: The values listed in this table are approximate. They were obtained using each enzyme's specific unit assay substrate DNA.
Buffers should be thawed completely and mixed thoroughly before using.
Unique NEBuffer Chart
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Restriction Endonucleases - NEB Online Catalog
Click for the Chart Legend.
|A|B|C|D |E|F|H|K |M|N|P|R |S|T|X|Z|
% Activity in NEBuffers
Enzyme
Supplied NEBuffer
1
2
3
4
Aat II
4
0
50
50
100
Acc I
4
50
50
10
100
Acc65 I
3 + BSA
10
75
100
25
Aci I
3
25
50
100
50
Acl I
4 + BSA
10
10
0
100
Acu I
2 + SAM
50
100
50
100
Afe I*
SE-Y
25
50
25
100
Afl II
2 + BSA
50
100
25
100
Afl III
3 + BSA
25
75
100
50
Age I
1
100
50
10
75
Ahd I
4 + BSA
25
75
0
100
Ale I
4
10
25
10
100
Alu I
2
100
100
75
100
Alw I
4
50
100
10
100
AlwN I
4
50
100
50
100
Apa I@25°C
4 + BSA
25
50
0
100
ApaL I
4 + BSA
100
100
10
100
ApeK I @75°C
3
25
75
100
50
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NEBuffer Activity Chart for Restriction Enzymes, New England Biolabs
Apo I@50°C
3 + BSA
10
75
100
75
Asc I
4
0
10
10
100
Ase I
3
NR
75
dd100
NR
AsiS I
3 + BSA
50
100
100
50
Ava I
4
10
75
10
100
Ava II
4
50
75
10
100
Avr II
2
100
100
50
100
Bae I@25°C
2 + BSA + SAM
50
100
50
75
BamH I
Udd + BSA
75
100
50
75
Ban I
4
50
100
50
100
Ban II
4
100
10
50
100
Bbs I
2
100
100
25
75
Bbv I
2
100
100
25
75
BbvC I
4
50
100
10
100
Bcc I
1 + BSA
100
50
10
50
BceA I
Udd + BSA
25
NR
NR
75
Bcg I
3dd + SAM
50
75
100
50
BciV I
4
100
25
10
100
Bcl I@50°C
3
50
100
dd100
75
Bfa I
4
75
50
10
100
BfrB I
4 + BSA
10
25
10
100
BfuA I@50°C
3
0
75
100
10
BfuC I
4 + BSA
100
50
25
100
Bgl I
3
50
75
100
50
Bgl II
3
10
75
100
10
Blp I
4
50
100
10
100
Bme1580 I
2
75
100
75
75
BmgB I
3 + BSA
0
25
100
10
Bmr I
2
75
100
75
100
Bmt I*
SE-W
25
100
10
50
Bpm I
3 + BSA
75
100
100
100
Bpu10 I*
SE-K
0
25
100
25
BpuE I@25°C
2 + SAM
75
100
10
75
Bsa I@50°C
3
75
75
100
100
BsaA I
3
75
75
100
75
BsaB I@60°C
2
75
100
100
100
BsaH I
4 + BSA
50
100
100
100
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NEBuffer Activity Chart for Restriction Enzymes, New England Biolabs
BsaJ I@60°C
2
100
100
100
100
BsaW I@60°C
2 + BSA
50
100
50
50
BsaX I
4
75
100
10
100
BseR I
2
100
100
75
100
BseY I
3
10
50
100
50
Bsg I
4 + SAM
50
75
50
100
BsiE I@60°C
2 + BSA
50
100
10
100
BsiHKA I@65°C
3 + BSA
50
100
100
100
BsiW I@55°C
3
100
100
100
25
Bsl I@55°C
3
10
50
100
75
Bsm I@65°C
2
75
100
75
75
BsmA I@55°C
3
50
100
100
50
BsmB I@55°C
3
75
100
100
100
BsmF I@65°C
4 + BSA
25
50
50
100
BsoB I
2
10
100
100
50
Bsp1286 I
4 + BSA
25
25
25
100
BspCN I@25°C
1 + BSA + SAM
100
75
10
75
BspD I
4
25
75
50
100
BspE I
3
0
10
100
0
BspH I
4
10
100
50
100
BspM I
3
NR
NR
100
NR
Bsr I@65°C
3
0
50
100
10
BsrB I
2
50
100
100
100
BsrD I@65°C
2 + BSA
50
100
50
75
BsrF I
U
25
100
75
75
BsrG I
2 + BSA
25
100
10
100
BssH II@50°C
3
100
100
dd100
100
BssK I@60°C
3 + BSA
0
50
100
50
BssS I
3
0
50
dd100
10
BstAP I* @60°C
SE-W
25
25
100
25
BstB I@65°C
4
75
50
25
100
BstE II@60°C
3 + BSA
50
75
100
75
BstF5 I* @65°C
SE-Y
75
50
25
100
BstN I@60°C
2 + BSA
10
100
100
75
BstU I@60°C
2
100
100
50
100
BstX I@55°C
3
25
100
100
50
BstY I@60°C
2
50
100
75
100
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NEBuffer Activity Chart for Restriction Enzymes, New England Biolabs
BstZ17 I
3
NR
NR
100
NR
Bsu36 I
3 + BSA
0
25
100
0
Btg I
3 + BSA
25
50
100
100
BtgZ I@60°C
4 + BSA
10
25
0
100
Bts I
4 + BSA
100
50
50
100
Cac8 I
3
50
75
100
75
Cla I
4 + BSA
10
50
50
100
CspC I
2
10
100
10
75
CviA II@25°C
4
75
25
10
100
Dde I
3
75
100
100
75
Dpn I
4
100
100
75
100
Dpn II
U
NR
NR
dd100
NR
Dra I
4
75
75
50
100
Dra III
3 + BSA
100
75
100
25
Drd I
4
25
50
10
100
Eae I
1
100
100
50
100
Eag I
3
10
25
100
10
Ear I
1
100
100
50
100
Eci I
2 + BSA
50
100
100
50
EcoN I
4
100
100
75
100
EcoO109 I
4 + BSA
100
100
75
100
EcoP15 I
3 + BSA + ATP
75
100
100
100
EcoR I
Udd
100
100
100
100
EcoR V
3 + BSA
50
75
100
50
Fat I* @55°C
SE-G
10
100
50
50
Fau I* @55°C
SE-B
100
25
10
50
Fnu4H I
4
10
25
25
100
Fok I
4
100
100
75
100
Fse I
4 + BSA
100
75
0
100
Fsp I
4
10
75
10
100
Hae II
4 + BSA
75
100
50
100
Hae III
2
50
100
25
75
Hga I
1
100
75
50
100
Hha I
4 + BSA
75
100
100
100
Hinc II
3 + BSA
75
100
100
100
Hind III
2
50
100
10
50
Hinf I
2
75
100
75
75
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NEBuffer Activity Chart for Restriction Enzymes, New England Biolabs
HinP1 I
2
100
100
100
100
Hpa I
4
25
50
10
100
Hpa II
1
100
50
10
100
Hph I
4
NR
75
0
100
Hpy188 I
4
50
25
10
100
Hpy188 III
4 + BSA
100
100
10
100
Hpy99 I
4 + BSA
10
10
0
100
HpyCH4 III
4
100
50
25
100
HpyCH4 IV
1
100
25
10
25
HpyCH4 V
4
50
50
25
100
I-Ceu I
4 + BSA
10
10
0
100
I-Sce I
U + BSA
10
50
50
50
Kas I
2 + BSA
75
100
75
75
Kpn I
1 + BSA
100
75
0
50
Mbo I
3
75
100
100
100
Mbo II
2
100
100
50
100
McrBC
2 + BSA + GTP
50
100
10
75
Mfe I
4
75
50
10
100
Mlu I
3
25
75
100
50
Mly I
4 + BSA
50
50
25
100
Mme I
4 + SAM
NR
NR
NR
dd100
Mnl I
2 + BSA
75
100
50
75
Msc I
4
75
75
75
100
Mse I
2 + BSA
75
100
75
75
Msl I
2
100
100
25
100
Msp I
2
75
100
50
50
MspA1 I
4 + BSA
50
100
50
100
Mwo I@60°C
3
10
75
100
75
N.Alw I
2
25
100
25
50
N.BbvC IA
2
50
100
10
100
N.BbvC IB
4
50
100
10
100
N.BstNB I @55°C
3
0
10
100
10
Nae I
1
100
75
10
75
Nar I
1
100
75
75
75
Nb.Bsm I @65°C
3
25
100
100
25
Nci I
4
100
25
10
100
Nco I
4
100
100
100
100
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NEBuffer Activity Chart for Restriction Enzymes, New England Biolabs
Nde I
4
75
100
75
100
NgoM IV
4
100
50
10
100
Nhe I
2 + BSA
100
100
10
100
Nla III
4 + BSA
25
25
25
100
Nla IV
4 + BSA
10
10
10
100
Not I
3 + BSA
0
50
100
25
Nru I
3
0
10
100
10
Nsi I
3
10
75
100
25
Nsp I
2 + BSA
100
100
0
100
Pac I
1 + BSA
100
75
10
100
PaeR7 I
4
25
100
10
100
Pci I*
SE-O
25
75
100
50
PflF I
4 + BSA
0
25
25
100
PflM I
3 + BSA
0
100
100
50
Pho I@75°C
3
50
50
100
75
PI-Psp I @65°C
U + BSA
0
10
10
10
PI-Sce I
U + BSA
0
0
0
0
Ple I
4
100
100
75
100
Pme I
4 + BSA
0
50
10
100
Pml I
1 + BSA
100
75
0
75
PpuM I
4
0
25
0
100
PshA I
4 + BSA
50
75
10
100
Psi I*
SE-B
50
100
0
25
PspG I@75°C
2
75
100
50
100
PspOM I*
SE-Y
25
100
50
100
PspX I*
SE-Y + BSA
10
100
10
50
Pst I
3 + BSA
75
75
100
50
Pvu I
3 + BSA
10
75
100
10
Pvu II
2
100
100
100
100
Rsa I
1
100
100
50
100
Rsr II
4
25
75
10
100
Sac I
1 + BSA
100
50
10
100
Sac II
4
25
75
10
100
Sal I
3 + BSA
0
0
100
0
Sap I
4
75
50
0
100
Sau3A I
U + BSA
100
50
10
100
Sau96 I
4
50
100
100
100
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NEBuffer Activity Chart for Restriction Enzymes, New England Biolabs
Sbf I
4
75
50
0
100
Sca I
3
100
100
dd100
25
ScrF I
4
100
100
100
100
SexA I
3 + BSA
25
25
100
100
SfaN I
3
0
75
100
25
Sfc I
4 + BSA
75
50
10
100
Sfi I@50°C
2 + BSA
0
100
10
100
Sfo I
2
25
100
50
50
SgrA I
4
100
50
10
100
Sma I@25°C
4
0
0
0
100
Sml I@55°C
4 + BSA
25
75
25
100
SnaB I
4 + BSA
25
50
25
dd100
Spe I
2 + BSA
75
100
25
75
Sph I
2
100
100
50
100
Ssp I
U
50
100
50
50
Stu I
2
100
100
50
100
Sty I
3 + BSA
25
75
100
10
StyD4 I
2
10
100
100
100
Swa I@25°C
3 + BSA
10
10
100
10
Taq I@65°C
3 + BSA
50
75
100
75
Tfi I@65°C
3
100
100
dd100
100
Tli I@75°C
3 + BSA
25
25
100
10
Tse I@65°C
3
75
100
100
75
Tsp45 I@65°C
1 + BSA
100
25
0
75
Tsp509 I@65°C
1
100
100
100
NR
TspR I@65°C
4 + BSA
25
50
25
100
Tth111 I@65°C
1
100
25
25
100
Xba I
2 + BSA
0
100
75
75
Xcm I
2
10
100
50
50
Xho I
2 + BSA
75
100
100
100
Xma I
4 + BSA
25
50
0
100
Xmn I
2 + BSA
100
100
75
100
Zra I*
SE-B
100
100
25
100
Click for the Chart Legend.
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