| Buffer Compatibility for Modifying Enzymes | - to print |
Fermentas 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 Fermentas enzymes and used in common applications.
* – 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.
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 Fermentas enzymes and used in common applications.
| DNA/RNA modifying enzyme | Enzyme activity in 1X buffers, % |
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| FastDigest® |
B |
G |
O |
R |
Tango 1X |
Tango 2X |
BamHI |
Ecl136II, SacI |
EcoRI |
KpnI |
Taq with KCl |
Taq with (NH4)2SO4 |
Pfu |
RT | T4 DNA Ligase |
|
| T4 DNA Ligase * |
75-100 | 100 | 100 | 75- 100 |
75- 100 |
75- 100 |
75- 100 |
75- 100 |
50 | 75- 100 |
100 | 75 | 75 | 75 | 75 | 100 |
| FastAP™ Thermosensitive Alkaline Phosphatase |
100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 50 | 75- 100 |
100 | nd |
| Shrimp Alkaline Phosphatase |
100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 75-100 | 100 | 100 | 100 | 100 | 100 | 100 | nd |
| T4 Polynucleotide Kinase ** |
100 | 75- 100 |
100 | 100 | 75- 100 |
100 | 100 | 100 | 50-75 | 100 | 75- 100 |
100 | 0 | 0 | 100 | 100 |
| DNA Polymerase I |
100 | 25-50 | 75- 100 |
100 | 100 | 100 | 100 | 100 | 50-75 | 100 | 50-75 | 100 | 100 | 100 | 100 | nd |
| Klenow Fragment |
100 | 25-50 | 20-50 | 100 | 100 | 100 | 100 | 100 | 50-75 | 100 | 50-75 | 100 | 100 | 100 | 100 | 100 |
| Klenow Fragment, exo- |
100 | 25-50 | 25-50 | 100 | 100 | 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 | 100 | 100 | 50 | 100 | 50 | 100 | 100 |
| T7 DNA Polymerase |
100 | 75- 100 |
100 | 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 | nd | nd | 100 | 100 | 100 | nd | nd |
| Exonuclease III | 0-25 | 100 | 25-50 | 0-25 | 0-25 | 25-50 | 0-25 | 0-25 | 100 | 0-25 | 100 | nd | nd | nd | nd | nd |
| Lambda Exonuclease |
nd | nd | 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 |
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 | 100 | 100 |
0-25 | 100 | 25-50 | 100 | 100 | 75-100 | 100 | 50-75 |
** – 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 Mgl2 |
0.1 mg/ml BSA |
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| G | 10 mM Tris-HCl (pH 7.5 at 37°C) |
10 mM Mgl2 | 50 mM NaCl |
0.1 mg/ml BSA | ||||
| O |
50 mM Tris-HCl (pH 7.5 at 37°C) |
10 mM Mgl2 | 100 mM NaCl |
0.1 mg/ml BSA | ||||
| R |
10 mM Tris-HCl (pH 8.5 at 37°C) |
10 mM Mgl2 | 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 | ||||
| 2X Tango™ |
66 mM Tris-acetate (pH 7.9 at 37°C) |
20 mM Mg-acetate |
132 mM K-acetate |
0.2 mg/ml BSA | ||||
| BamHI |
10 mM Tris-HCl (pH 8.0 at 37°C) |
5 mM Mgl2 | 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 Mgl2 | 0.1 mg/ml BSA | |||||
| EcoRI |
50 mM Tris-HCl (pH 7.5 at 37°C) |
10 mM Mgl2 | 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 Mgl2 | 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 Mgl2 | 50 mM KCl |
0.08% Nonidet P40 |
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| Taq with (NH4)2SO4 |
75 mM Tris-HCl (pH 8.8 at 25°C) |
2 mM Mgl2 | 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 Mgl2 | 50 mM KCl |
10 mM DTT |
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| T4 DNA Ligase |
140 mM Tris-HCl (pH 7.8 at 25°C) |
10 mM Mgl2 | 0.5 mM ATP |
10 mM DTT |




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