2× PCR Super Mix HYA441
2× PCR Super Mix is an optimized premix containing DNA polymerase, dNTPs, MgCl2 , KCl and other stabilizers. It only needs to add primers and templates to perform amplification. This product can use complex genomic DNA as a template to amplify a target fragment of 5 kb in length or a simple template such as lambda DNA to amplify a target fragment of 10 kb in length. It is suitable for conventional PCR reaction, vector construction, and other applications.
Components
|
Components |
Size-1 |
Size-2 |
Size-3 |
|
2× PCR Super Mix |
5mL |
1 mL × 15 |
1 mL × 100 |
Properties
|
Specification |
Function |
|
5´-3´exonuclease activity |
No |
|
3´-5´exonuclease activity |
Yes |
|
Fidelity |
6X Taq |
|
Product End |
Blunt end |
Storage
Store at -20°C.
Instructions
1. Reaction System
|
Components |
25uL Reaction System |
50uL Reaction System |
|
2× PCR 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 ℃ |
45 s* |
1 |
|
Denaturation |
98 ℃ |
10 s |
30 |
|
Annealing |
55-65 ℃ |
30 s |
|
|
Extension |
72℃ |
20-30 s/kb |
|
|
Final Extension |
72℃ |
5 min |
1 |
|
Hold |
4-12 ℃ |
- |
1 |
*Note: In the case of high complexity DNA templates, the pre-denaturation time should be extended up to 3 minutes for fully denaturation.