Hot Start Taq DNA Polymerase (Glycerol-Free)
Croyez Hot Start Taq DNA Polymerase contains Taq DNA Polymerase and an anti-taq monoclonal antibody which blocks polymerase activity. Enzyme activity is recovered during the initial incubation step while the taq antibody is denatured and dissociates from the DNA polymerase. Hot Start Taq DNA Polymerase exhibits higher specificity, sensitivity, and yield by reducing non-specific amplification and primer-dimers. This enzyme possesses 5’→3’ polymerase activity and 5’→3’ exonuclease replacement activity, but lacks a 3’→5’ exonuclease (proof-reading) activity. Hot Start Taq DNA Polymerase is suitable for most PCR applications.
The enzyme formulation does not contain glycerol and is compatible for further lyophilization process.
The enzyme formulation does not contain glycerol and is compatible for further lyophilization process.
Package & Component:
Purity:
>98% as determined by SDS-PAGE analysis.
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C.
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Shipping Conditions:
Blue ice
| Cat. | Name | Amount |
| C15030-500U | Hot Start Taq DNA Polymerase (Glycerol-Free) (5 U/μL) | 500 U |
| 10X Hot Start Taq Buffer (Mg2+ free) | 1 mL | |
| 25 mM MgCl2 | 1 mL | |
| C15030-1000U | Hot Start Taq DNA Polymerase (Glycerol-Free) (5 U/μL) | 1000 U |
| 10X Hot Start Taq Buffer (Mg2+ free) | 2 X 1 mL | |
| 25 mM MgCl2 | 2 X 1 mL |
Purity:
>98% as determined by SDS-PAGE analysis.
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 74°C.
Storage:
Stored at -20°C. Avoid repeated freeze/thaw cycles.
Shipping Conditions:
Blue ice
The following procedure is a general guideline for qPCR reaction. To maintain an RNase-free environment, always wear disposable gloves, and use laboratory consumables and water of nuclease-free grade during the whole experiment course.
PCR reaction set-up:
1. Place all required reagents on ice.
*See Usage Notes for additional guidelines on primer/template preparation.
2. Gently mix the reaction thoroughly to achieve uniform distribution and briefly centrifuge.
3.Thermal cycling conditions for standard PCR.
PCR reaction set-up:
1. Place all required reagents on ice.
| Component | Amount | Final concentration |
| 10X Hot Start Taq Buffer (Mg2+ free) | 5 μL | 1 X |
| 10 mM dNTPs | 1 μL | 0.2 mM each |
| Forward primer (10 μM) | 1-5 μL | 0.2-1 μM |
| Reverse primer (10 μM) | 1-5 μL | 0.2-1 μM |
| 25 mM MgCl2 | 2-8 μL | 1-4 μM |
| Template DNA | X μL | ≦1 μg |
| Hot Start Taq DNA Polymerase (Glycerol-Free) | 0.25 – 0.5 μL | 1.25 – 2.5 U |
| Nuclease-Free H2O | Y μL | - |
| Total reaction volume | 50 μL | - |
2. Gently mix the reaction thoroughly to achieve uniform distribution and briefly centrifuge.
3.Thermal cycling conditions for standard PCR.
| Step | Cycles | Temperature | Time |
| Initial denaturation / Enzyme activation | 1 | 95°C | 5 min |
| Denaturation | 35-50 | 95°C | 10–20 sec |
| Annealing/Extension | 60°C | 20–60 sec |
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