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Description
A new generation of polymerase that delivers improved yield, sensitivity, speed and robustness when amplifying targets from any template.
Product Highlights
- Sensitive – incorporates MyTaq DNA polymerase that exhibits increased affinity for DNA thereby improving amplification of even limiting amounts of template
- Efficient – novel buffer system maximizes efficiency of PCR amplification, delivering improved yield of any PCR product
- Robust – reliable amplification in the presence of inhibitors and with even the most challenging DNA targets
- Flexible – ideal for amplifying any target up to 5 kb from DNA extracted from human, animal and plant samples
- Convenient – buffer system includes a red dye for improved pipetting ease / accuracy and to enable direct gel loading
- Fast – developed to give sensitive, reproducible and robust amplification of a broader range of targets under fast thermal cycling conditions
Product Description
MyTaq™ Red DNA Polymerase is recommended for all standard PCR applications. The MyTaq DNA Polymerase and MyTaq Red Reaction Buffer in this product, are a unique combination of next-generation polymerase and novel buffer system that deliver very high yield PCR amplification over a wide range of PCR templates. MyTaq has an increased affinity for DNA, enabling reliable amplification from even very low amounts of template. MyTaq DNA Polymerase has been developed to give more robust amplification than other commonly-used polymerases allowing it to perform well with challenging templates and in the presence of PCR inhibitors. Furthermore, the highly efficient nature of MyTaq means it gives excellent results under fast PCR conditions.
The enzyme is supplied with a 5x MyTaq Red Reaction Buffer that increases the visual contrast between the reagent and the reaction vessel for improved convenience and to improve pipetting accuracy. The red dye also enables samples to be loaded directly on to a gel after the PCR without the need to add loading buffer. In addition, the MyTaq Red Reaction Buffer contains dNTPs, MgCl2 and enhancers at optimal concentrations, which helps eliminate the need for optimization, thereby saving time, effort and the cost of performing unnecessary assay repeats.
Applications
- Standard PCR
- High-yield PCR
- Fast PCR
- Colony PCR
- TA cloning
Highly efficient and robust amplification under a broad range of PCR conditions.
MyTaq DNA Polymerases Customer Review
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 researchApplication Note
MyTaq™ Red DNA PolymeraseWe use MyTaq Red DNA Polymerase weekly in the lab for our mouse genotyping. Fast, efficient, reliable and cost effective. Thanks Bioline!
Nora Tenis, SVI, Molecular Genetics, Fitzroy, AU
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 |
500 Units |
2500 Units |
5000 Units |
|
MyTaq Red DNA Polymerase |
1 x 100 µL |
2 x 250 µL |
4 x 250 µL |
|
5x MyTaq Reaction Buffer |
4 x 1 mL |
14 x 1.5 mL |
9 x 5 mL |
Concentration
5 u/μL
Storage & Stability
MyTaq Red DNA Polymerase can be stored for up to 6 months at -20°C, or up to 2 weeks at +4°C. Repeated freeze/thaw cycles should be avoided. When stored under the recommended conditions and handled correctly, full activity of the kit is retained until the expiry date printed on the outer box label.
Shipping conditions
On Dry Ice or Blue Ice.
FAQs
Yes, MyTaq DNA Polymerase does not possess 3' to 5' exonuclease activity, and therefore leaves an A-overhang at the 3' end, thus making the PCR product suitable for integration into TA cloning vectors.
The MyTaq HS DNA polymerase works very well with difficult samples like GC rich or bisulfite converted DNA. Especially the hot-start is important, as it decreases primer dimer and unspecific amplification. We would suggest to start with the recommended protocol. If optimization is needed, we would suggest to optimize the annealing temperature. Due to the degraded DNA after bisulfite conversion it may be useful to increase the amount of template and polymerase.
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 polymerase and primer can be increased, but do not exceed the suggested limits.
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.
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.
The MyTaq HS DNA polymerase works very well with difficult samples like GC rich or bisulfite converted DNA. Especially the hot-start is important, as it decreases primer dimer and unspecific amplification. We would suggest to start with the recommended protocol. If optimization is needed, we would suggest to optimize the annealing temperature. Due to the degraded DNA after bisulfite conversion it may be useful to increase the amount of template and polymerase.
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.
No, the dyes and composition of the Red Reaction Buffers and Mixes are such that the samples will sink easily into the well and the samples can be clearly seen, thus no loading buffer is required to load your samples on an agarose gel.
The dye present in the Red Mixes or Red Reaction Buffers does not interfere with PCR product isolation procedures or subsequent applications like sequencing. An exception is the quantification of PCR products in colored buffers with photometric methods or fluorescence assays. We cannot exclude the impact of the dye on quantification results and suggest the purification of these samples, for instance with ISOLATE II PCR & Gel kit or SureClean Plus.
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