2× PCR Premix （without Dye） HYA443
PCR Master Mix is a kind of conventional PCR premixed solution which is ready to use, including Taq DNA Polymerase, dNTP mix MgCl2 and optimized buffer. During the reaction, only the primer and template can be added for amplification, which greatly simplifies the operation steps of experiment. This product contains excellent stabilizers and can be stored for 3 months at 4℃. The PCR product have 3'-dA protrusion and can be easily cloned in to T vector.
Components
|
Components |
Size-1 |
Size-2 |
Size-3 |
|
2× PCR Premix (without Dye) |
5mL |
1 mL×15 |
1 mL×50 |
Storage
Store at -20°C for 2 years.
Instructions
1. Reaction System
|
Components |
Volume |
|
2× PCR Premix (without Dye) |
25 uL |
|
Forward Primer (10 μM) |
2 µL |
|
Reverse Primer (10 μM) |
2 µL |
|
DNA Template* |
Variable |
|
ddH2O |
Up to 50 µL |
*Template usage: 50-200 ng genomic DNA; 0.1-10 ng plasmid DNA
2. Reaction Program
|
Step |
Temperature |
Time |
Cycles |
|
Initial Denaturation |
94 ℃ |
5 min |
1 |
|
Denaturation |
94 ℃ |
30 s |
35 |
|
Annealing |
50-60 ℃ |
30 s |
|
|
Extension |
72 ℃ |
30-60 sec/kb |
|
|
Final Extension |
72 ℃ |
10 min |
1 |
|
Hold |
4 ℃ |
- |
1 |
1. Mg2+ concentration: This product contains 3 mM of MgCl2, suitable for most PCR reactions.
2. Annealing temperature: Please refer to the theoretical Tm value of primers. The annealing temperature can be set to 2-5℃ lower than the theoretical value of the primer.
3. Extention time: For molecular identification, 30 sec/kb is recommended. For gene cloning, 60 sec/kb is recommended.
Notes
1. For your safety and health, please wear lab coats and disposable gloves for operation.
2. It is used for research use ONLY!