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 |
MegaFi™ Pro Fidelity 2X PCR MasterMix sets a new benchmark for specific, sensitive, robust, and high-fidelity PCR performance in a uniquely-formaulated buffer with gel loading dye. This advanced, strategically engineered enzyme is combined with a monoclonal antibody that inhibits both 5’-3’ polymerase activity and 3'-5’ exonuclease activity at room temperature, enabling precise Hot Start reactions. With exceptional sensitivity, MegaFi™ Pro can amplify even the most challenging templates, outperforming leading competitors. It delivers unparalleled high fidelity with an ultra-low error rate—over 2,000 times lower than Taq polymerase—making it the most accurate enzyme available. Ideal for demanding applications such as Next Generation Sequencing, molecular cloning, and diagnostics, MegaFi™ Pro ensures the highest quality results for your PCR needs. Product Features:
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| Application |
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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? | |
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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? | |
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Blunt ends, this makes it not suitable for TA cloning
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| What does 3' - 5' exonuclease activity mean? | |
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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? | |
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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? | |
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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? | |
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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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| Can I store my PCR reaction mix once it has been assembled? | |
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It is generally not recommended to store an assembled PCR reaction for long periods, as the activity of the polymerase can decrease over time. However, you can prepare the reaction mix and store it on ice for short periods (a few hours) before running the PCR. Always prepare fresh reactions when possible for the best results.
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| What discontinued abm product is Cat. No. G887 equivalent to? | |
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This product is abm’s next generation of DNA Polymerases and is functionally equivalent to Cat. G124, G124-dye, G464, G464-dye, G497, G497-dye, G886, and G897 (Precision, Bestaq, Kodaq, and MegaFi PCR MasterMixes), with improved performance. Contact our customer service team technical@abmgood.com for more information.
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| What is the source of this enzyme? | |
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The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.
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