2× HiFi Super Mix HYA447
2× HiFi Super Mix is an ideal product with good amplification efficiency for diversity templates including animals, plants, cDNA, etc. 2× HiFi Super Mix contains HS DNA polymerase , dNTP, and an optimized buffer system. It only needs to add primers and templates to perform amplification, thereby reducing pipetting operations, significantly controlling cross-contamination among samples, and improving detection throughput and reproducibility of results.The amplification system contains protective agents that keep 2× HiFi Super Mix stable in activity after repeated freezing and thawing.
Components
|
Components |
Size-1 |
Size-2 |
|
2× HiFi Super Mix |
1 mL |
1 mL×5 |
Properties
|
5´-3´exonuclease activity |
No |
|
3´-5´exonuclease activity |
Yes |
|
Fidelity |
100X Taq |
|
Product End |
Blunt end |
Storage
Store at -20°C.
Instructions
1. Reaction System
|
Components |
25uL Reaction System |
50uL Reaction System |
|
2× HiFi Super Mix |
12.5uL |
25uL |
|
Forward Primer (10 μM) |
0.5 µL |
1 µL |
|
Reverse Primer (10 μM) |
0.5 µL |
1 µL |
|
DNA Template* |
Variable |
Variable |
|
Nuclease-free Water |
Up to 25 µL |
Up to 50 µL |
*Note: The optimal reaction concentration varies with different DNA templates. Please refer to the basic principles of PCR below.
2. Reaction Program
|
Step |
Temperature |
Time |
Cycles |
|
Initial Denaturation |
98 ℃ |
3 min |
1 |
|
Denaturation |
98 ℃ |
10 s |
25-35 |
|
Annealing |
55-65 ℃ |
20-30 s |
|
|
Extension |
72℃ |
30-60 s/kb* |
|
|
Final Extension |
72℃ |
1-5 min |
1 |
|
Hold |
4-12 ℃ |
- |
1 |
*Properly extending the extension time can improve the amplification yield. For complex amplification templates, such as genomic DNA, it is recommended to extend at a speed of 60 s/kb, and more recommended conditions please refer to the basic principles of PCR below.