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Description
Unique blend of highly-efficient MyTaq HS DNA Polymerase and a proprietary proofreading enzyme that combine to give increased target affinity for use with challenging templates and inhibitor-rich samples.
Product Highlights
- Robust - enzyme blend and buffering system promotes reliable amplification of the most challenging and complex targets, even in the presence of inhibitors
- Sensitive - improved target affinity and high processivity ensure successful amplification in low-copy number assays
- Efficient - high-yield amplification of a broad range of targets up to 10 kb including complex DNA extracted from human, animal and plant samples
- Specific - an antibody-mediated hot-start blend that remains completely inactive during PCR set-up to prevent non-specific amplification
- Convenient - an all-in-one-tube mastermix that improves the speed, convenience and accuracy of PCR set-up
- Accurate - proofreading component delivers 3.5x higher fidelity than Taq DNA Polymerase, enabling cloning of PCR products
Product Description
MyFi™ has been developed to give reliable amplification of targets up to 10 kb from challenging and complex targets. MyFi shows improved tolerance to PCR inhibitors, thereby enabling reliable detection from samples from which DNA is difficult to purify. Furthermore a unique buffer system and enzyme blend promote highly sensitive amplification, ideal for low-copy number targets. The proofreading ability of MyFi allows all PCR products to be cloned. The inclusion of MyTaq HS means MyFi Mix generates PCR products with 3’-A overhangs, which is perfect for TA cloning. MyFi Mix has the added convenience of room temperature reaction assembly, to avoid non-specific amplification and primer-dimer formation.
MyFi Mix is supplied as a mastermix that requires the addition of only template, primers and water, thereby reducing the number of pipetting steps during PCR set-up, for improved speed, throughput and assay reproducibility. The inclusion of dNTPs, MgCl2 and enhancers at optimal concentrations, helps eliminate the need for optimization, thereby saving on time, cost and making MyFi Mix well suited to automation.
Applications
- Amplification of challenging and complex templates
- Inhibitor-tolerant PCR
- Longer PCR (up to 10 kb)
- Robust PCR
- Low-copy PCR
- Higher-fidelity PCR (NGS library amplification)
- TA cloning
High-yield PCR from GC-rich and complex templates.
PCR Enzyme Guide
Download the PCR Enzyme Guide with detailed product descriptions and performance data to help you choose the best product for your researchPCR Selection Chart
Select the best reagent for your researchMyFi Mix is more sensitive than our current Taq and so easy to use/set up. Nice product.
Aberystwyth University, IBERS, UK
Product Selection
Please refer to the PCR Selection Chart to confirm the recommended product for your PCR application.Resources
Documents
Certification of Analysis (COAs)
Specification
Components
|
Reagent |
100 Reactions |
500 Reactions |
|
MyFi Mix, 2x |
2 x 1.25 mL |
10 x 1.25 mL |
Volume
- BIO-25049: 100 x 50 µL Reactions: 2 x 1.25 mL
- BIO-25050: 500 x 50 µL Reactions: 10 x 1.25 mL
Concentration
- 2x
Storage & Stability
All components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided.
When stored under the recommended conditions and handled correctly, full activity is retained until the expiry date indicated on the outer box label.
Shipping conditions
Shipped on Dry Ice or Blue Ice.
FAQs
At Bioline we pride ourselves in supplying high-quality polymerases to suit your requirements. To aid your selection of the most suited enzyme for your specific applications, please see our enzyme selection tool.
All our polymerases are guaranteed for a period of 12 months from the date of purchase. These should be stored at -20°C during this time for optimal retention of activity.
Please Note: We do not recommend the storage of our polymerases at -80 °C as ice crystals could form on the active site, which may affect or destroy the activity of the enzyme.
Please Note: We do not recommend the storage of our polymerases at -80 °C as ice crystals could form on the active site, which may affect or destroy the activity of the enzyme.
PCR can be a challenging technique, with various parameters to optimize to achieve the best results. If you are having problems, these could be easily resolved by addressing a few issues. Please see our PCR troubleshooting guide for suggestions and help with your specific problems.
| Observation | Recommended Solution(s) |
| No or low PCR yield | Enzyme concentration too low – increase the amount of enzyme in 0.5 U increments. |
| Primers degraded – check quality and age of the primers. | |
| Magnesium concentration too low – increase concentration in 0.25 mM increments with a starting concentration of 1.75 mM. | |
| Primer concentration not optimized. Titrate primer concentration (0.3-1 µM); ensuring that both primers have the same concentration. | |
| Template concentration too low – Increase concentration of template. | |
| Perform a positive control to ensure that the enzyme, dNTPs and buffers are not degraded and/or contaminated. | |
| Multiple Bands | Primer annealing temperature too low. Increase annealing temperature. Primer annealing should be at least 5°C below the calculated Tm of primers. |
| Prepare master mixes on ice or use a heat-activated polymerase. | |
| For problems with low specificity. Try adding 3% DMSO (not supplied) to improve specificity. | |
| Smearing or artifacts | Template concentration too high. Prepare serial dilutions of template. |
| Too many cycles. Reduce the cycle number by 3-5 to remove non-specific bands. | |
| Enzyme concentration too high - decrease the amount of enzyme in 0.5 U increments. | |
| Extension time too long. Reduce extension time in 0.5-1 minute increments. |
These terms refer to parameters to be considered when performing PCR and are important features in selecting the correct enzyme for your needs. Understanding what they mean is therefore crucial:
Yield: The amount of DNA produced in a PCR reaction.
Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand.
Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified.
Specificity: A measure of the unwanted by-products generated in a reaction.
During PCR setup at room temperature, standard polymerases will have some activity. When using a hot-start polymerase, this enzyme will have no activity at room temperature, thus reducing the risk of unspecific products in the reaction due to these mis-primed oligonucleotides. This feature makes this type of polymerase ideally suited to high-throughput applications, where the reactions may be left sitting at room temperature for prolonged periods of time.
In nature, the ability of a DNA polymerase to correct misincorporations of nucleotides in the DNA strand being elongated is often crucial to the survival of the host organism. This ability is termed "proofreading activity" and occurs in the 3' to 5' direction. This activity also leads to the polymerase removing unpaired nucleotides overhanging at the 3'end (A-overhangs), creating blunt ends.
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 minutes at 72°C.
All Bioline polymerases are available in the convenient format of a 2x master mix, containing all the reagents and additives necessary to perform successful PCR, with the exception of template, primers and water.
This formulation not only provides a convenient and hassle-free PCR setup, but also significantly reduces the chances of human error, inaccuracies and contamination by reducing the pipetting steps required.
We recommend a final reaction volume of 50 µL, this requiring the use of 25 µL of the 2x master mix and to make up to 50 µL using template, primers and PCR grade water.
Whilst the exact composition of the mixes is proprietary information, all mixes are made up of reaction buffer, magnesium, dNTPs, polymerase and additives, designed to keep the polymerase stable in the mix, as well as provide the optimal conditions for PCR.
All Bioline PCR mixes come supplied at a 2x concentration, requiring the user simply to add template, primers and PCR grade water to a final concentration of 1x.
My DNA sample contains PCR inhibitors, which impairs my PCR results. What can I do for optimization?
If possible, we would recommend an additional cleanup of affected samples, for instance, customers had very good experiences with the clean-up of samples containing humic acids with SureClean Plus. Alternatively decreasing the template concentration will help to minimize the amount of inhibitors. The amount of primer can be increased, but do not exceed the suggested limits.
Please click here in order to request your sample. You will receive an email confirmation within two business days with delivery details.
