2× PCR Premix （with Dye） HYA442
2× PCR Master Mix contains Taq DNA Polymerase, dNTPs, and other PCR-required components. The Master Mix is stable for 3 months at 4℃ with our customized stabilizers. The pre-mix solution is optimized for conventional PCR and ready to use by adding DNA template and primers. The PCR products can be loaded directly for electrophoresis with pre-loaded bromophenol blue dye. The amplified products contain 3 '-dA protrusion and can be easily cloned into T vector. The 2× PCR Master Mix simplifies PCR procedure and reduces contamination.
Components
|
Components |
Size-1 |
Size-2 |
Size-3 |
|
2× PCR Premix (with 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 (with 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. PCR products with 2× PCR Master Mix are not suitable for polyacrylamide gel electrophoresis.
2. For your safety and health, please wear lab coats and disposable gloves for operation.
3. It is used for research use ONLY!