Restriction Enzyme Activity In Promega Buffers
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1
2Restriction Enzyme Activity in Promega 10X Buffers, Reaction
Temperature and Heat Inactivation
Restriction Enzyme Activity in Promega 10X Buffers, Reaction Temperature and Heat Inactivation.
The 10X Reaction Buffer supplied with each restriction enzyme is optimized to give 100% activity. In many cases, good activity is also obtained using one of the
4-CORE
®
10X Buffers. Many commonly used cloning enzymes have Buffers E and H as their optimal buffer, and so we have determined the activity of many of our
enzymes in these buffers. This table may be used to select the best buffer for digestion with multiple restriction enzymes. Alternatively, visit the Restriction Enzyme
Resource at: Enzyme activity is expressed as a percent of the activity obtained with the optimal buffer for each enzyme.
Buffer
Activity in Buffer
Enzyme
Promega
Supplied
MULTI-
Heat
Assay
Enzyme
with Enzyme
A
B
C
D
E
H
CORE™
Inactivation Temperature
AatII
J
50–75%
10–25%
<10%
<10%
10–25%
<10%
<10%
+
37°C
AccI
G
50–75%
25–50%
25–50%
10–25%
<10%
<10%
25–50%
–
37°C
AccIII
F
<10%
10–25%
25–50%
25–50%
n.d.
n.d.
<10%
–
65°C
100%
100–125%**
100%
Acc65I
D
10–25%
50–75%
75–100%
75–100%
+
37°C
AccB7I
E
10–25%
50–75%
100%*
<10%
100%
n.d.
100%
+
37°C
100%
AgeI
K
25–50%
25–50%
25–50%
50–75%
n.d.
n.d.
+
37°C
AluI
B
75–100%
100%
75–100%
10–25%
n.d.
n.d.
10–25%
+
37°C
100%
100%
Alw44I
C
<10%
25–50%
25–50%
n.d.
n.d.
+
37°C
ApaI
A
100%
50–75%
50–75%
<10%
10–25%
<10%
75–100%
+
37°C
100%
100%
AvaI
B
10–25%
50–75%
25–50%
10–25%
<10%
+/–
37°C
AvaII
C
50–75%
50–75%
100%
25–50%
n.d.
n.d.
25–50%
+
37°C
BalI
G
10–25%
<10%
<10%
<10%
n.d.
n.d.
<10%
+
37°C
BamHI
E
75–100%*
75–100%
75–100%
50–75%
100%
50–75%
75–100%
+
37°C
BanI
G
25–50%
25–50%
10–25%
<10%
n.d.
n.d.
100%
–
50°C
100%
BanII
E
75–100%
75–100%
75–100%
25–50%
n.d.
n.d.
+
37°C
BbuI
A
100%
75–100%
75–100%
<10%
10–25%
10–25%
100%
+
37°C
BclI
100%
C
10–25%
75–100%
50–75%
50–75%
50–75%
10–25%
–
50°C
BglI
D
10–25%
25–50%
75–100%
100%
25–50%
75–100%
100%
+
37°C
100%
BglII
D
25–50%
75–100%
75–100%
n.d.
n.d.
<10%
–
37°C
BsaMI
D
10–25%
25–50%
50–75%
100%
n.d.
n.d.
25–50%
–
65°C
100%
Bsp1286I
A
50–75%
25–50%
10–25%
n.d.
n.d.
75–100%
+
37°C
BsrSI
D
10–25%
25–50%
10–25%
100%
n.d.
n.d.
100%
–
65°C
100%
BssHII
H
75–100%
50–75%
75–100%
50–75%
n.d.
75–100%
–
50°C
Bst98I
D
<10%
10–25%
10–25%
100%
n.d.
n.d.
25–50%
–
37°C
100%
100%
BstEII
D
25–50%
50–75%
50–75%
n.d.
n.d.
–
60°C
BstOI
C
10–25%
25–50%
100%
25–50%
n.d.
n.d.
<10%
–
60°C
BstXI
D
<10%
10–25%
25–50%
100%
100%
75–100%
10–25%
+/–
50°C
BstZI
100%
D
<10%
<10%
10–25%
10–25%
75–100%
10–25%
–
50°C
Bsu36I
E
<10%
25–50%
50–75%
25–50%
100%
n.d.
50–75%
–
37°C
100%
100%
CfoI
B
75–100%
75–100%
25–50%
n.d.
n.d.
+/–
37°C
ClaI
C
75–100%
75–100%
100%
75–100%
100%
50–75%
100%
+
37°C
CspI
100%
100–125%**
K
<10%
10–25%
25–50%
50–75%
10–25%
+
30°C
Csp45I
B
25–50%
100%
50–75%
25–50%
100%
25–50%
50–75%
+
37°C
100%
DdeI
D
25–50%
25–50%
50–75%
n.d.
n.d.
25–50%
+/–
37°C
DpnI
B
50–75%
100%
75–100%
50–75%
n.d.
n.d.
100%
+
37°C
100%
DraI
B
75–100%
75–100%
50–75%
n.d.
n.d.
25–50%
+
37°C
EclHKI
E
<10%
<10%
75–100%
10–25%
100%
n.d.
50–75%
+
37°C
100%
Eco47III
D
<10%
25–50%
50–75%
n.d.
n.d.
25–50%
+
37°C
EcoICRI
B
10–25%
100%
75–100%
<10%
25–50%
n.d.
100%
+
37°C
EcoRI
H
25–50%
50–75%
50–75%
50–75%
75–100%
100%
100%*
+
37°C
100%
100%
EcoRV
D
10–25%
25–50%
50–75%
25–50%
50–75%
+
37°C
HaeII
B
50–75%
100%
50–75%
10–25%
n.d.
n.d.
100%
–
37°C
100%
100%
HaeIII
C
75–100%
75–100%
50–75%
n.d.
n.d.
–
37°C
HhaI
C
50–75%
75–100%
100%
50–75%
n.d.
n.d.
75–100%
+
37°C
100%
100%
HincII
B
25–50%
25–50%
50–75%
75–100%
50–75%
+
37°C
HindIII
E
25–50%
100%
75–100%
10–25%
100%
25–50%
50–75%
+
37°C
100%
HinfI
B
50–75%
75–100%
75–100%
n.d.
n.d.
50–75%
–
37°C
HpaI
J
25–50%
50–75%
25–50%
10–25%
n.d.
n.d.
100%
–
37°C
100%
100%
HpaII
A
50–75%
50–75%
10–25%
n.d.
n.d.
–
37°C
(continued on next page)
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