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 | FastDigest (Green), Tango 1X and Tango 2X | B | G | O | R | BamHI | Ecl136II, SacI | EcoRI | KpnI | Taq with KCl | Taq with (NH4)2SO4 | Pfu | RT | T4 DNA Ligase |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Enzyme activity in 1X buffers, % | ||||||||||||||
| T4 DNA Ligase* | 75-100 | 100 | 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 | 50 | 75- 100 | 100 | nd |
| T4 Polynucleotide Kinase (T4 PNK)** | 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 |
** 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 | ||||
| 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.