2 × Phanta Flash Master Mix
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.
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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. 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 inhibiting the 5'→3' polymerase activity and 3'→5' exonuclease activity at room temperature, which enable 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. Amplification will generate blunt-ended products, which are compatible with ClonExpress kits (Vazyme #C112/C113/C115) and TOPO cloning kit (Vazyme #C601).
Applications
This product is suitable for PCR with various templates such as genomic DNA, cDNA, plasmid DNA, dU-containing DNA, crude samples, etc.
Notes
- For fragments ≤10 kb, the recommended extension time is 4 - 5 sec/kb. For fragments >10 kb, the recommended extension time is 10 sec/kb.
- High quality templates should be used to ensure successful amplification and products yield.
- Phanta Flash Super-Fidelity DNA Polymerase has strong proof-reading activity. If TA cloning needs to be performed, please perform purification before dA-tailing.
- Primer Design
- It is recommend that the last base at the 3' end of primer should be G or C.
- Consecutive mismatches should be avoided in the last 8 bases at the 3' end of the primer.
- Avoid hairpin structures at the 3' end of the primer.
- Differences in the Tm value of the forward primer and the reverse primer should be no more than 1℃ and the Tm value should be adjusted to 55℃ to 65℃ (Primer Premier 5 is recommended to calculate the Tm value).
