Description 2 × Phanta Flash Master Mix is a new generation superior enzyme based on Phanta Flash Super-Fidelity DNA Polymerase. Through directed optimization of Phanta DNA Polymerase, Phanta Flash Super-Fidelity DNA Polymerase has the characteristics of rapid amplification (4 - 5 sec/kb) while maintaining high fidelity and yield. The amplification error rate of Phanta Flash Super-Fidelity DNA Polymerase is 81-fold lower than that of conventional Taq DNA Polymerase. Matched with optimized buffer system, this kit can achieve high amplification specificity. And it has excellent compatibility with crude samples, templates with uracil and GC-rich system (primer/template). This kit contains two types of monoclonal antibodies that inhibit the 5'→3' polymerase activity and 3'→5' exonuclease activity at room temperature, enabling it to perform hot start PCR with great specificity. It contains all required reaction components (Phanta Flash Super-Fidelity DNA Polymerase, dNTP and optimized buffer), except primers and templates, thereby simplifying the operation process and improving the detection throughput and repeatability. This product contains tracking dyes, so PCR products can be directly loaded for electrophoresis after the reaction. Features High fidelity: ~81X higher than Taq DNA Polymerase Flash amplification: Extension speed up to 5 sec/kb Robust performance: Superior specificity, sensitivity, and yields Broad Compatibility: Suitable for complex samples & GC-rich systems Components Components P520-01 P520-02 P520-03 2 × Phanta Flash Master Mix (Dye Plus) 1ml 5×1ml 15×1ml Experimental Cases Ultra-fast Amplification 2 × Phanta Flash Master Mix (Vazyme#P510), 2 × Phanta Flash Master Mix (Dye Plus) (Vazyme#P520) and other commercially available brands of high-fidelity polymerases were used to perform PCR amplification. The target fragment of 353 bp and 873 bp were amplified at an extension time of 1 sec/kb, 7140 bp fragment was amplified at an extension time of 4 sec/kb and 5 sec/kb, respectively. The results show that all the fragments could be successfully amplified by P510 and P520. Ultra-fast Amplification Amplification of fragments ≤10 kb can be achieved within 1 h! High Specificity Vazyme#P510, Vazyme#P520 and other commercially available brands of high-fidelity polymerases were used to amplify different length of fragments in accordance to the recommended systems and procedures of respective instructions. The results show that P510 and P520 have higher amplification specificity than other brands of high-fidelity polymerases. Storage Store at -30 ~ -15°C and transport at ≤0°C. FAQ Q1: Does the tracking dye in 2 × Phanta Flash Master Mix (Dye Plus) interfere with downstream applications? A1: The tracking dye (which consists of Bromophenol Blue) is compatible with downstream applications such as fluorescent automated DNA sequencing, ligation, and restriction digestion. For applications that require analysis of PCR products by absorbance or fluorescence excitation, we recommend using colorless versions of the products Phanta Flash Super-Fidelity DNA Polymerase or 2 × Phanta Flash Master Mix or purifying the PCR products prior to analysis. Q2: Can I perform restriction digestion in the Phanta Flash reaction buffer? A2: 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. Q3: Can I amplify GC-rich targets with Phanta Flash Super-Fidelity DNA Polymerase? A3: 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. Q4: What are your recommendations when working with long amplicons using Phanta Flash Super-Fidelity DNA Polymerase? A4: 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. Q5: What are your recommendations when calculating annealing temperatures using Phanta Flash Super-Fidelity DNA Polymerase?A5: Approximately 5°C above Tm of primers, but no more than 68°C. Product Manual Manual_P520 V23.1 Purchase Help pcr buffer,master mix
