HotStart PCR Master Mix is a ready-to-use, conventional PCR premix solution containing Hot Start Taq DNA Polymerase, dNTP mixture, MgCl2 and an optimized buffer system. The reaction can be carried out by simply adding primers and templates, which greatly simplifies the experimental procedure. The product contains bromophenol blue dye, and the PCR product can be directly electrophoresed. This product contains excellent stabilizers and can beplacedfor3months at 4 °C. The PCR product has a 3′-dA overhang and can be easily cloned into a T vector.
| Reaction System | |||
| Components | Volume（μL） | ||
| Template DNA | Moderate amount | ||
| Primer 1(10 μM) | 2 | ||
| Primer 2(10 μM) | 2 | ||
| HotStart PCR MasterMix | 25 | ||
| ddH2O | to 50 | ||
| Amplification Procedure | |||
| Cycle step | Temperature（℃） | Time | Cyclenumber |
| Pre-denaturation | 94 | 5 min | 1 |
| Denaturation | 94 | 30 sec | 35 |
| Annealing | 50-60 | 30 sec | |
| Extended | 72 | 30 sec/kb | |
| Final Extended | 72 | 10 min | 1 |