DNA Safe Stains
Hot start Taq DNA polymerase
Description
Hot start Taq DNA Polymerase is designed for Real-Time PCR and Hot- start PCR. It is modified with a special inhibition of PCR at room temperature. This will prevent primer dimers and other artifacts.
Storage buffer
50mM Tris-HCl pH7.9, 50mM KCl, 0.1mM EDTA,
1mM DTT, 0.5mM PMSF , 50% glycerol.
10X reaction buffer
Buffer containing 25mM MgCl2
Unit description
One unit is defined as the amount of enzyme that will incorporate 10nmole of dNTP into acid-insoluble material in 30 minutes at 74ºC. The reaction conditions are: 50mM Tris-HCl pH8.8, 50mM NaCl, 5mM MgCl2, 200uM each of dATP, dCTP, dGTP, dTTP, 10 mg activated calf thymus DNA and 0.1mg/ml BSA in a final volume of 50 ml.
Source
E coli clone
Genomic DNA: 10–200 ng
Plasmid DNA : 1–5 ng
cDNA : ~100 ng starting total RNA
Amplification of longer targets (up to 15 kb) may be possible, but may require more template and longer elongation times.
Applications
– Hot Start and real time PCR
– Multiplex PCR
– Amplification of complex genomic and cDNA
| Cat No | Pack size | Conc. |
| TAMB02Z-500 | 500U | 5U/uL |
| TAMB02Z-1000 | 1000U | 5U/uL |
| TAMB03Z-2500 | 2500U | 5U/uL |
| PCR cycles program | |||
|---|---|---|---|
| Step | Temperature | Time | Cycle |
| Initial denaturation | 94-95°C | 10 mins | 1 |
| Denaturation | 94-95°C | 10-60sec | 25-35 |
| Annealing | 50-68°C | 10-30sec | |
| Extension | 72°C | 1min/1kb | |
| Final extension | 72°C | 1-10 mins | 1 |
| IIMPORTANT: Annealing temperature should be 2-6ºC lower than the primer melting temperature. |
|---|
Shipping and Storage conditions
Shipping and temporary storage at -20 and for up to 1 month at room temperature has no detrimental effects on the quality of Hot start DNA polymerase.
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