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Restriction Enzyme Buffers


Fermentas


Five Buffer System


NEB

1X Buffer B (blue)
10 mM Tris-HCl (pH 7.5 at 37°C),
10 mM MgCl2,
0.1 mg/ml BSA
1X Buffer SacI:
10 mM Bis-Tris Propane-HCl (pH 6.5 at 37°C)
10 mM MgCl2 and 0.1 mg/ml BSA. 		1X NEBuffer 1:
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.0 @ 25°C
1X Buffer G (green)
10 mM Tris-HCl (pH 7.5 at 37°C), 
10 mM MgCl2, 
50 mM NaCl, 
0.1  mg/ml BSA 		1X NEBuffer 2:
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C
1X Buffer O (orange)
50 mM Tris-HCl (pH 7.5 at 37°C), 
10 mM MgCl2, 
100 mM NaCl, 
0.1 mg/ml BSA 		1X NEBuffer 3:
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C
1X Buffer R (red)
10 mM Tris-HCl (pH 8.5 at 37°C), 
10 mM MgCl2, 
100 mM KCl, 
0.1 mg/ml BSA 		1X NEBuffer 4:
50 mM potassium acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM Dithiothreitol
pH 7.9 @ 25°C
1X Buffer Tango™ (yellow)
33 mM Tris-acetate (pH 7.9 at 37°C), 
10 mM magnesium acetate, 
66 mM potassium acetate,
0.1  mg/ml BSA

Promega

MULTI-CORE™ Buffer (1X):
25mM Tris-Acetate
pH 7.5 (at 37°C)
100mM potassium acetate
10mM magnesium acetate
1mM DTT.
1X Buffer for BamHI
10 mM Tris-HCl (pH 8.0 at 37°C), 
5 mM MgCl2, 
100 mM KCl,
0.02% Triton X-100,
1 mM 2-mercapthoethanol,
0.1 mg/ml BSA 		1X NEBuffer 3:
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C
1X Buffer for EcoRI
50 mM Tris-HCl (pH 7.5 at 37°C), 
10 mM MgCl2, 
100 mM NaCl,
0.02% Triton X-100,
0.1 mg/ml BSA 		1X NEBuffer EcoRI:
100 mM Tris-HCl
50 mM NaCl
10 mM MgCl2
0.025 % Triton X-100
pH 7.5 @ 25°C


Promega 4-CORE® Buffer System

Restriction Enzymes Resource

Composition of Promega Restriction Enzyme Reaction Buffers (1X)

Buffer

pH
(at 37°C)

Tris-HCl
(mM)

MgCl2
(mM)

NaCl
(mM)

KCl
(mM)

DTT
(mM)

  A

7.5

6

6

6

-

1

  B 

7.5

6

6

50

-

1

  C

7.9

10

10

50

-

1

  D

7.9

6

6

150

-

1

  E

7.5

6

6

100

-

1

  F

8.5

10

10

100

-

1

  G

8.2

50

5

-

-

-

  H

7.5

90

10

50

-

-

  J

7.5

10

7

-

50

1

  K

7.4

10

10

-

150

-

  L

9.0

10

3

100

-

-

MULTI-CORE™ Buffer (1X) = 25mM Tris-Acetate, pH 7.5 (at 37°C), 100mM potassium acetate, 10mM magnesium acetate, 1mM DTT.

Notes:

  1. For each 10°C rise in temperature between 0°C and 25°C, the pH of Tris buffers decreases 0.31 pH units.
  2. For each 10°C rise in temperature between 25°C and 37°C, the pH of Tris buffers decreases 0.25 pH units.

Pharmacia

One-Phor-All Buffer PLUS

This buffer is originally from Pharmacia (SE) and now available from GE Healthcare

Recipe (10x): (Source)

  • 1M potasssium glutamate
  • 250 mM tris-acetate
  • 100 mM magnesium acetate
  • pH 7.5

One-Phor-All Buffer PLUS activity

(Source)
One-Phor-All Buffer PLUS activity guide

The table that follows is a guide to the activity of some of our restriction enzymes in One-Phor-All Buffer PLUS (OPA+). Enzymes not listed in the table have not been tested for activity in OPA+ buffer. For each enzyme in the table the following pieces of information are provided:

The optimum concentration of OPA+ buffer - the concentration at which the enzyme's activity is highest (the buffer is a 10× concentrate1).
The enzyme's activity in the optimum concentration of OPA+ buffer as a percentage of its activity in the assay buffer listed on the Certificate of Analysis.

Percentages are designated as follows: ++ = 50-99%; +++ = 100%; ++++ = 101-199%; +++++ > 200%

Restriction enzyme

Optimum OPA buffer conc.

Activity in OPA as a percentage of activity in assay buffer

Aat II

1×

+++++

Acc I

1×

++

Alu I

1×

+++

Apa I

0.5×

+++++

Ava I

2×

+++

Ava II

1×

+++++

Bam H I

1×

++++

Bgl I2

1×

++

Bgl II

1×

+++

Cla I

1×

+++++

Dde I

1×

+++++

Dra I

1×

+++

EcoR I3

1×

+++

EcoR V

2×

+++

Hae III

1×

+++

Hha I

1×

+++

Hinc II

1×

+++

Hind III

1×

+++

Hinf I

1×

+++++

Hpa I

1×

+++

Hpa II

1×

+++

Kpn I

1×

+++

Mlu I

2×

+++

Msp I

1×

+++

Nar I

1×

+++

Nco I 3

1×

++++

Nde I

2×

++

Nhe I

1×

+++

Not I4

2×

++++

Pst I

1×

+++++

Pvu I

2×

++++

Pvu II5

1×

+++

Rsa I

1×

+++++

Sac I

1×

+++

Sac II

1×

++

Sal I

2×

++

Sau3A I

1×

+++

Sca I

2×

+++++

Sfi I

1×

+++

Sma I

1×

+++

Sph I

1×

+++

Stu I

1×

+++

Taq I

1×

++++

Xba I

1×

+++

Xho I

1×

+++++

1   OPA+ buffer (27-0901-02) will be provided free of charge upon request. Please inquire.
2   With Bgl I, addition of KCl to a final concentration of 100 mM KCl is required.
3   With EcoR I and Nco I, star activity may be observed in 1× OPA+ when DNA is digested for longer than 1 hour. Star activity can be eliminated during extended incubations by using 2× concentration.
4   With Not I, addition of Triton X-100 to a final concentration of 0.01% (w/v) is required.
5   With Pvu II, addition of NaCl to a final concentration of 50 mM is required.