Description
Hieff Unicon Hotstart Direct Taq DNA Polymerase is a hotstart DNA polymerase that is resistant to blood and other inhibitors. The product is blocked by antibodies and has good amplification sensitivity and specificity. After heating at the pre-denaturation temperature, the antibody is completely inactivated and the DNA polymerase activity is released. The amplification induced by nonspecific annealing of primers could be effectively inhibited by using the hotstart Taq enzyme.
Components
|
Components No. |
Name |
10717ES72 (250 U) |
10717ES80 (1000 U) |
10717ES92 (10 KU) |
10717ES93 (25 KU) |
10717ES94 (50 KU) |
|
10717 |
Hotstart D-Taq (5 U/μL) |
50 μL |
200 μL |
1 mL× 2 |
1 mL× 5 |
10 mL |
Storage
The product should be stored at -20°C for 2 years.
Instructions
1. Reaction Composition
|
Components |
Volume (μL) |
Final Concentration |
|
2 × Buffer |
25 |
1 × |
|
Primer/Probe Mix |
X |
0.1 -0.5 μM |
|
Hotstart D-Taq (5 U/μL) |
1.2 |
0.12 U/μL |
|
Template DNA |
X |
0.1-100 ng |
|
ddH2O |
to 50 |
- |
Note：Be sure to mix well before use, avoid excessive bubbles caused by violent vibration.
a) According to the specific experimental application, it is needed to prepare the corresponding reaction buffer. If a basic buffer (10×) is required, Cat#11374 is recommended.
b) The amount of DNA and the concentration of probes or primers are recommended concentrations. The optimal concentration can be adjusted according to the specific experimental conditions.
c) The amount of enzyme can be adjusted according to the experimental application.
2. Optimized Cycling Protocol
|
|
Reaction stage |
Temperature |
Time |
Cycle |
|
1 |
Initial denaturation |
95°C |
5 min |
1 |
|
2 |
Amplification reaction |
95°C |
15 sec |
45 cycles |
|
60°Ca |
30 secb |
Note：
a) Amplification reaction: The temperature is adjusted according to the Tm value of the designed primers.
b) Fluorescence signal acquisition: Please set the experimental procedure according to the requirements of the instrument manual.