Modifying Enzyme Buffers
Thermo Scientific DNA/RNA modifying enzymes are supplied with optimized reaction buffers.
However, it is often convenient to use these enzymes in other buffers for experiments that involve multiple enzymatic reactions. The table below lists activities of DNA/RNA modifying enzymes in common reaction buffers, supplied with Thermo Scientific Molecular Biology enzymes and used in common applications.
DNA/RNA modifying enzyme | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Tango 2X |
Taq with KCl | |||||||||||||
T4 DNA Ligase * |
75-100 | 100 | 100 | 75- 100 |
75- 100 |
75- 100 |
50 | 75- 100 |
100 | 75 | 75 | 75 | 75 | 100 |
FastAP
Thermo- sensitive Alkaline Phosphatase |
100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 50 | 75- 100 |
100 | nd |
T4 Polynucleotide Kinase
** |
100 | 75- 100 |
100 | 100 | 75- 100 |
100 | 50-75 | 100 | 75- 100 |
100 | 0 | 0 | 100 | 100 |
DNA Polymerase I |
100 | 25-50 | 75- 100 |
100 | 100 | 100 | 50-75 | 100 | 50-75 | 100 | 100 | 100 | 100 | nd |
Klenow Fragment |
100 | 25-50 | 20-50 | 100 | 100 | 100 | 50-75 | 100 | 50-75 | 100 | 100 | 100 | 100 | 100 |
Klenow Fragment, exo-
|
100 | 25-50 | 25-50 | 100 | 100 | 100 | 50-75 | 100 | 75- 100 |
100 | 100 | 100 | 100 | nd |
T4 DNA Polymerase |
100 | 75- 100 |
75- 100 |
100 | 100 | 100 | 100 | 100 | 100 | 50 | 100 | 50 | 100 | 100 |
T7 DNA Polymerase |
100 | 75- 100 |
100 | 100 | 100 | 75- 100 |
75-100 | 100 | 100 | nd | nd | nd | nd | nd |
Exonuclease I | nd | nd | nd | nd | nd | nd | nd | nd | nd | 100 | 100 | 100 | nd | nd |
Exonuclease III | 0-25 (FastDigest (Green), Tango 2X) 25-50 (Tango 1X) |
100 | 25-50 | 0-25 | 0-25 | 0-25 | 100 | 0-25 | 100 | nd | nd | nd | nd | nd |
Lambda Exonuclease |
nd | nd | nd | nd | nd | nd | nd | nd | nd | 50-75 | 50-75 | 50- 75 |
nd | nd |
phi29 DNA
Polymerase |
100 | 25-100 | 100 | 100 | 100 | 100 | 50-75 | 100 | 50-75 | 25-50 | 75-100 | 75-100 | 75-100 | 75-100 |
Bsm
DNA Polymerase |
100 | 25-50 | 75-100 | 100 | 100 | 100 | 0-25 | 100 | 25-50 | 100 | 100 | 75-100 | 100 | 50-75 |
*
Buffers were supplemented with 0.5 mM ATP, which is required for T4 DNA
Ligase activity.
**The
activity of this enzyme was compared to its activity in buffer A (for
forward reaction).
nd – not determined.
1X Buffer Composition
B | 10 mM Tris-HCl (pH 7.5 at 37°C) |
10 mM MgCl2 |
0.1 mg/mL BSA |
|||||
G | 10 mM Tris-HCl (pH 7.5 at 37°C) |
10 mM MgCl2 | 50 mM NaCl |
0.1 mg/mL BSA | ||||
O |
50 mM Tris-HCl (pH 7.5 at 37°C) |
10 mM MgCl2 | 100 mM NaCl |
0.1 mg/mL BSA | ||||
R |
10 mM Tris-HCl (pH 8.5 at 37°C) |
10 mM MgCl2 | 100 mM KCl |
0.1 mg/mL BSA | ||||
Tango |
33 mM Tris-acetate (pH 7.9 at 37°C) |
10 mM Mg-acetate |
66 mM K-acetate |
0.1 mg/mL BSA | ||||
BamHI |
10 mM Tris-HCl (pH 8.0 at 37°C) |
5 mM MgCl2 | 100 mM KCl |
0.02% Triton X-100 |
0.1 mg/mL BSA | 1 mM BME |
||
Ecl136II, SacI |
10 mM Bis-Tris Propane-HCl (pH 6.5 at 37°C) |
10 mM MgCl2 | 0.1 mg/mL BSA | |||||
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 | |||
KpnI |
10 mM Tris-HCl (pH 7.5 at 37°C) |
10 mM MgCl2 | 0.02% Triton X-100 |
0.1 mg/mL BSA | ||||
Taq with KCl |
10 mM Tris-HCl (pH 8.8 at 25°C) |
1.5 mM MgCl2 | 50 mM KCl |
0.08% Nonidet P40 |
||||
Taq with (NH4)2SO4 |
75 mM Tris-HCl (pH 8.8 at 25°C) |
2 mM MgCl2 | 0.01% Tween 20 |
20 mM (NH4)2SO4 | ||||
Pfu |
20 mM Tris-HCl (pH 8.8 at 25°C) |
2 mM MgSO4 | 10 mM KCl |
0.1% Triton X-100 |
0.1 mg/mL BSA | 10 mM (NH4)2SO4 |
||
RT |
50 mM Tris-HCl (pH 8.3 at 25°C) |
4 mM MgCl2 | 50 mM KCl |
10 mM DTT |
||||
T4 DNA Ligase |
140 mM Tris-HCl (pH 7.8 at 25°C) |
10 mM MgCl2 | 0.5 mM ATP |
10 mM DTT |
DNA/RNA Modifying Enzyme Dilution
DNA/RNA modifying enzymes are supplied in their optimal storage buffers which are specially formulated for long term storage. If required for specific applications, dilute the enzyme with 1X reaction buffer for short term use.
For Research Use Only. Not for use in diagnostic procedures.