p-Superhot Taq DNA Polymerase: Datasheet
Features: Maximo p-Superhot Taq DNA Polymerase for qPCR is designed for Real Time PCR. Polyclonal antibodies inhibit the reaction at room temperature until after the first denaturation step. This prevents primer-dimers and other artefacts. Using the enzyme there is no need to adjust the existing standard PCR protocol in your lab.
Applications:
- Hot Start and real time PCR
- Multiplex PCR
- Amplification of complex genomic and cDNA templates
- no primer-dimers and other artefacts; inactive at room temperature
- short activation time
- enhanced PCR sensitivity
Description:
Maximo p-Superhot Taq DNA is an optimized mixture of a high processive Taq DNA Polymerase and polyclonal antibodies to Taq DNA. The enzyme is a thermostable DNA polymerase that possesses a 5´→3´ polymerase activity and a double-strand specific 5´→3´ exonuclease activity. The enzyme consists of a single polypeptide with a molecular weight of 94kDa.
Concentration: 5 u/µl
Unit definition:
One unit incorporates 10 nmol of deoxyribonucleotide into acid-precipitation material in 30 min at 72°C degree
Storage Buffer:
50 mM Tris-HCl, pH 8.2, 0.1 mM EDTA, 0.1% Tween 20, 0.1% NP40, 1 mM DTT, 50% Glycerol
Reaction Buffer:
10X Buffer: 100mM KCl, 80mM (NH4)2SO4, 100mM Tris-HCl, pH9.0, 0.5% Nonident P40, 15 mM MgSO4
Quality control:
Activity and performance test, SDS-PAGE purity, absence of endonucleases/nickases and exonucleases test
Usage:
Use your existing and optimized protocol. In contrast to chemically modified Taq DNA pol. where the first denaturation step needs up to 15 min, Maximo p-Superhot Taq does not need a prolonged heating or denaturation time.
|
Components |
Volume per reaction |
|
10X reaction buffer |
5 µl |
|
Mg+ |
optional |
|
dNTP-Mix (40mM) |
1.0 µl |
|
Up-stream primer (10 µM stock) |
0,5-2.5 µl |
|
Down-stream primer (10µM stock) |
0.5-2,5 µl |
|
Template DNA |
0.1-15 ng/ml plasmid DNA
1-10 µg/ml genomic DNA |
|
Maximo p-Superhot Taq DNA (5 u/µl) |
0.2 - 1.0 µl |
|
Sterile dest. Water (molecular bio. grade) |
up to 50 µl total reaction volume |
Note:
- vortex all solutions carefully before using
- add the enzyme after Template DNA
- may you have to optimize the MgCl2 concentration for best result
General Thermo-Cycler protocol:
|
Step |
Time |
Temperature |
|
Initial denaturation |
4-5 min |
94-95°C |
25-30 Cycles:
Denaturation
Annealing
Extension
|
10-25 sec
10-25 sec
60 sec |
94-95°C
55-65°C
72°C per 1kb
|
|
Final extension |
5 min |
72°C |
Note:
- In case of low amount of DNA template, additionally cycles may be used
Transportation: on blue ice
Storage: at -20°C for 24 months
|
