The polymerase chain reaction (PCR) is a technique for producing significant amounts of specific DNA from a limited number of complex templates as well as detecting DNA.1, 2
PCR uses the property that DNA polymerase can replicate DNA sequentially from the 5' end to the 3' end according to the principle of complementary base pairing.3
The 2 × Phanta Flash Master Mix includes the Phanta Flash Super-Fidelity DNA Polymerase, a new generation superior enzyme with excellent rapid amplification performance (≤1 kb fragment, 1 sec/kb; ≤10 kb fragment, 4 to 5 sec/kb; >10 kb fragment, 10 sec/kb).
This kit, when combined with an optimized buffer system, can achieve high amplification specificity and is compatible with crude samples (fungi, bacteria, blood samples, animal/plant tissue lysis products, and so on), uracil-based templates and GC-rich systems.
The 2 × Phanta Flash Master Mix includes all reaction components excluding primers and templates, making the operation process easier.
As the 2 × Phanta Flash Master Mix (Dye Plus) contains an electrophoresis indicator, there is no need for additional loading buffer after the PCR reaction is complete, and the samples can be spotted for electrophoresis right away.
Amplification of fragments smaller than 10 kb can be completed in an hour.
Optimized buffer system for higher specificity.
PCR products can be loaded directly for electrophoresis without the need for a loading buffer.
To perform PCR amplification, 2 × Phanta Flash Master Mix (Vazyme #P510), 2 × Phanta Flash Master Mix (Dye Plus) (Vazyme #P520), and other high-fidelity enzymes (TB1 and TB2) were used. Human genomic DNA and Arabidopsis genomic DNA were used as templates.
The 353 bp and 873 bp target fragments were amplified at a rate of 1 sec/kb, while the 7140 bp fragment was amplified at 4 sec/kb and 5 sec/kb, respectively.
The reaction system and experimental procedures were performed according to the product manual’s recommendations. The results in Figure 1 reveal that P510 and P520 were able to successfully amplify all of the fragments.
Figure 1. Comparison of amplification efficiency of P510 & P520 and other commercially available high-fidelity enzymes (TB1 and TB2). Image Credit: Vazyme Biotech Co. Ltd.
P510 and P520, as well as other commercially available high-fidelity enzymes, were used to amplify templates of mouse genomic DNA, human genomic DNA and Arabidopsis genomic DNA. The amount of DNA template utilized in each of them was fixed at 10 ng.
The reaction system and experimental procedures were performed according to the product manual’s recommendations. In comparison to other brands of high-fidelity enzymes, P510 and P520 have considerable yield and specificity advantages, according to the test results.
Figure 2. Comparison of yield and specificity of P510 & P520 and other commercially available high-fidelity enzymes. Image Credit: Vazyme Biotech Co. Ltd.
- Schochetman G, Ou C Y, Jones W K. Polymerase chain reaction[J]. The Journal of infectious diseases, 1988, 158(6): 1154–1157.
- DOES W P C R. Polymerase chain reaction[J]. Journal of Investigative Dermatology, 2013, 133.
- Joshi M, Deshpande J D. Polymerase chain reaction: methods, principles and application[J]. International Journal of Biomedical Research, 2010, 2(1): 81–97.
About Vazyme Biotech Co. Ltd.
Since the establishment in 2012, Vazyme has been dedicated to our mission "Science and Technology Make a Healthier Life" to focus on technology innovation and continuously expand the application fields of core technologies in life science, bio-medicine, and in vitro diagnostics.
As a R&D based company, we have been holding ourselves to the highest standards of ethics, accountability and professionalism. Our global research and development operations make sure we could provide quality products, solutions, and services locally to our customers, and more importantly, to do as much as it can to meet the unmet customers’ needs. For now, we are present in more than 60 countries and regions worldwide to get close to local customers.
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