2× Fast PCR Master Mix (with Dye)
Based on 2 publication(s) in Google Scholar
MCE 2× Fast PCR Master Mix (with Dye) is provided as a simple-to-use, stabilized 2× formulation that includes all components for fast PCR except sample DNA, primers and water, in which bromophenol blue dye is included. The 1 mL is defined as the base specification. All larger sizes correspond to incremental volumes of this base.
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Storage :
Store at -20°C for 2 years. 4°C for short-term storage (up to 3 months). Avoid repetitive freeze-thaw cycles while using.
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Description & Advantages
MCE 2× Fast PCR Master Mix (with Dye) is a ready-to-use premix for rapid PCR amplification. The product contains modified Taq DNA Polymerase, extension-enhancement factor, dNTPs Mix, MgCl2 and optimized reaction buffer. The amplification speed is up to 15 sec/kb and the maximum amplification speed is 5 sec/kb within 1 kb. With the primers and template added, the optimized system will provide sensitive and reliable DNA synthesis. After the PCR reaction, the PCR products can be loaded directly onto an agarose gel. It saves preparation time, reduces the risk of contamination from multiple pipetting steps, and provides consistent reaction-to-reaction performance. The PCR products contain poly(A) at the 3'-end and can be directly cloned into T-Vectors.
Protocol
1. Completely thaw and gently mix all components. Prepare the PCR reaction on ice as follows:
| Component | 50 μL reaction |
|---|---|
| 2× Fast PCR Master Mix (with Dye) | 25 μL |
| Forward Primer (10 μM) | 2 μL |
| Reverse Primer (10 μM) | 2 μL |
| DNA | X μL |
| ddH2O | To 50 μL |
Note: A final primer concentration of approximately 0.4 μM for each primer is recommended. Optimize template amount, annealing temperature, and cycle number according to the template and target fragment.
2. Recommended fast PCR program:
| Step | Temperature | Time | Cycles |
|---|---|---|---|
| Initial denaturation | 95°C | 3 min | 1 |
| Denaturation | 95°C | 10 sec | 30-35 |
| Annealing | 50-65°C | 10 sec | 30-35 |
| Extension | 72°C | 10-15 sec/kb | 30-35 |
| Final extension | 72°C | 5 min | 1 |
3. Bromophenol blue dye is included in the Master Mix, so PCR products may be loaded directly onto an agarose gel after amplification.
Publications
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Journal Impact Factor
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Most Recent
Storage
Store at -20°C for 2 years. 4°C for short-term storage (up to 3 months). Avoid repetitive freeze-thaw cycles while using.
Components
| Components | HY-K0532- 1 mL | HY-K0532- 5 mL | HY-K0532- 20 mL |
|---|---|---|---|
| 2× Fast PCR Master Mix (with Dye) | 1 mL | 1 mL × 5 | (1 mL × 5) × 4 |
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