Product Description
2 × Phanta Flash Master Mix utilizes the new generation of Phanta Flash Super-Fidelity DNA Polymerase. Through the directed evolution of Phanta DNA Polymerase, Phanta Flash Super-Fidelity DNA Polymerase has the characteristics of high fidelity (mismatch rate is 1/106 of conventional Taq DNA polymerase) and rapid amplification (5 sec/kb). With the application of the latest hot-start technology and the addition of specific factors, its amplification specificity and success rate are significantly improved. 2 × Phanta Flash Master Mix contains Phanta Flash Super-Fidelity DNA Polymerase, dNTP and optimized buffer. It only requires the addition of primers and templates for amplification, thereby reducing pipetting operations andimproving detection throughput and reproducibility of results. The amplification products of 2 × Phanta Flash Master Mix are blunt-ended, which are subsequently applicable for ClonExpress Ultra One Step Cloning Kit V2 (Vazyme #C116) and Ultra-Universal TOPO Cloning Kit (Vazyme #C603).
Features
High fidelity: ~106X higher than Taq DNA Polymerase
Flash amplification: Extension time up to 5 sec/kb
Robust performance: Superior specificity, sensitivity, and yields
Broad compatibility: Suitable for complex samples & GC-rich systems
Experimental Cases
Components
Storage
Store at -30 ~ -15°C and transport at ≤0°C.
FAQ
Q1: Can I perform restriction digestion in the Phanta Flash reaction buffer?
A1: Phanta Flash reaction buffer is compatible with restriction digestion directly after PCR. Note that Phanta Flash Super-Fidelity DNA Polymerase is not inactivated during PCR, thus purification of PCR products before restriction digestion is recommended if intact 5'- or 3'- overhangs are needed for cloning.
Q2: Can I amplify GC-rich targets with Phanta Flash Super-Fidelity DNA Polymerase?
A2: Phanta Flash Super-Fidelity DNA Polymerase can amplify targets with high GC content (up to 83% GC) without any additional DNA melting agents. In cases of extremely GC-rich targets (>83% GC), addition of DMSO to final concentration of 5% is recommended.
Q3: What are your recommendations when working with long amplicons using Phanta Flash Super-Fidelity DNA Polymerase?
A3: Good lab practices are important for long fragment amplification. These include using high-quality templates (pure, fresh, and intact) and fresh primer solutions. Reducing primer concentration to 0.3 µM may also improve the results.
Q4: What are your recommendations when calculating annealing temperatures using Phanta Flash Super-Fidelity DNA Polymerase?
A4: Approximately 5°C above Tm of primers, but no more than 68°C.
