MegaFi™ Pro Fidelity 2X PCR MasterMix
Cat. No. | G887 | ||||
Name | MegaFi™ Pro Fidelity 2X PCR MasterMix | ||||
Unit | 800 rxn (10.0 ml) | ||||
Category | PCR Polymerase | ||||
Description |
2000X lower error rates, 3X more affordable!MegaFi™ Pro Fidelity 2X PCR MasterMix elevates the current standards for specific, sensitive, robust, and high-fidelity PCR performance in a uniquely-formulated buffer with gel loading dye. This strategically-engineered polymerase is partnered with a monoclonal antibody which inhibits 5’ → 3’ polymerase activity and 3’ → 5’ exonuclease activity at room temperature allowing for highly specific Hot Start reactions. It has exceptional sensitivity and can amplify even the most difficult templates (compared to the leading competitors). Furthermore, MegaFi™ Pro boasts of its high fidelity and ultralow error rates (over 2,000X less than Taq polymerase, representing the lowest error rate on the market), making it incredibly useful for a variety of PCR applications demanding high fidelity, including Next Generation Sequencing, molecular cloning, or diagnostics. Product Features:
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Concentration | 2X | ||||
Storage Condition |
Store at -20°C. This product is stable for 2 years from the date of shipping if stored and handled properly. |
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Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G887 |
What are the advantages of using a High-Fidelity DNA Polymerase? | |
MegaFi Pro has high fidelity and high processivity. This means it has low error rates leading to more accurate sequences and longer amplicons leading to a faster reaction time. High fidelity is achieved due to its strong proofreading exonuclease activity, making it the preferred polymerase for all PCR applications requiring greater accuracy or the amplification of difficult templates.
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What ends will my PCR products have? | |
Blunt ends, this makes it not suitable for TA cloning
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What does 3' - 5' exonuclease activity mean? | |
It is the proofreading activity which digests nucleotides with 3´ hydroxyl groups from the 3´ to 5´ direction. This includes correction of mismatched base pairs and terminal 3´ digestion.
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Does the dye in the MasterMix affect downstream applications? | |
The loading dye does not interfere with downstream cloning experiments.
However, PCR clean-up is still advised prior to cloning due to the presence of excess primers, salts, and other buffer components that could potentially affect cloning efficiency.
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How do you determine the fidelity of MegaFi™ Pro? | |
The fidelity of MegaFi Pro has been tested through Blue-white screening assay.
LacZa gene was amplified using MegaFi™ Pro, NEB's Q5, Vazyme's PhantaMax, and Native Taq. The PCR amplicons were then subcloned into pUC19 vector and plated into a X-gal containing agar plate. Any mutations from PCR will cause the colonies to turn while no mutation will remain blue. Blue and white colonies were counted and the error rate per base was calculated and compared to native Taq.
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I am observing positive results in my negative controls (no template added) when attempting to detect gene sequences in a bacterial strain. I suspect this may be due to residual bacterial DNA from the DNA polymerase. What steps should I take to address this issue? | |
The presence of residual bacterial DNA is quite common in commercially available DNA polymerases, as most are expressed and purified using E. coli recombinant systems. In this case, we recommend switching to our Ultra-Pure BlasTaq™ 2X PCR MasterMix (Cat. No. G885). This product undergoes a rigorous multi-step purification protocol utilizing physical, chemical, and enzymatic methods to maximize the removal of contaminating genomic DNA.
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