Hifi HS DNA Polymerase Ⅰ HYD614
Hifi HS DNA Polymerase Ⅰ is a high-fidelity, thermostable DNA polymerase obtained through directed evolution of enzyme engineering. It has extremely strong 3’ to 5’ exonuclease activity and extremely low mismatch rate during PCR. Through specific antibody modification, it has no 5’ to 3’ polymerase activity and 3’ to 5’ exonuclease activity at room temperature, and can perform highly specific hot start PCR. The buffer contains extension auxiliary factors to maintain the stability and efficiency of the amplification process, and can easily realize conventional gene cloning, long fragment amplification and other experiments.
Components
|
Components |
100U |
500U |
1000U |
|
Hifi HS DNA Polymerase Ⅰ（1000U/mL） |
100μL |
500μL |
500μL×2 |
|
5×HiFi Buffer Ⅰ |
1000μL |
1000μL×5 |
1000μL×10 |
Storage
Store at -20℃. Mix well before use and avoid repeated freezing and thawing.
Instructions
1. Preparation of PCR reaction system
|
Composition |
Volume |
|
Template |
X µL |
|
Primer1 (10µM) |
2 µL |
|
Primer2 (10µM) |
2 µL |
|
5×HiFi Buffer Ⅰ |
10 µL |
|
Hifi HS DNA Polymerase Ⅰ |
1 |
|
ddH2O |
Up to 50 |
2. PCR amplification procedure
|
Step |
Temperature |
Time |
Cycles |
|
Initial denaturation |
95 ℃ |
3 mins |
1 |
|
Denaturation |
98 ℃ |
10 s |
25-35 |
|
Annealing |
60 ℃* |
15 s |
|
|
Extension |
72 ℃ |
1min/Kb** |
|
|
Fully extended |
72 ℃ |
5 min |
1 |
|
Hold |
4 ℃ |
∞ |
1 |
*Adjust the annealing temperature according to the primer Tm value, and the recommended setting is 60°C.
**When amplifying fragments below 10Kb, set it to 1min/Kb. When amplifying fragments of 10~20Kb, set the total extension time to 10min.
Notes
1. This product is for scientific research purposes only.
2. Please use high-quality templates.
3. Hifi HS DNA Polymerase Ⅰ is modified with monoclonal antibodies. The reaction system can be configured at room temperature, but the enzyme and buffer should be placed on ice before use and stored at -20℃ after use.
4. Primer design must meet general principles. If the 5' end has a modified sequence, that is, a sequence that is not paired with the template, it will not be included in the Tm value calculation.