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Description
MangoTaq™ DNA Polymerase is a formulation of Taq DNA Polymerase that offers high-yield across a wide range of DNA concentrations.
MangoTaq DNA Polymerase possesses 5´-3´ exonuclease activity and leaves an ´A´ overhang, resulting in PCR product suitable for effective integration into TA cloning vectors.
Product Highlights
Direct gel loading - no need for further post-PCR processing steps
Easy visual recognition- reduces pipetting errors
Robust performance - perfect for a wide range of PCR reactions
MangoTaq™ DNA Polymerase offers high yield across a wide range of DNA concentrations. MangoTaq DNA Polymerase leaves an ´A´ overhang such that the PCR product is suitable for effective integration into TA cloning vectors.
The polymerase is supplied with two different reaction buffers for greater flexibility. For high-throughput applications, MangoTaq and the colored reaction buffer make an ideal choice, since this combination enables the user to load directly on a gel in order to facilitate easy recognition.
The two reaction buffers supplied are: 5x Colored Reaction Buffer and 5x Colorless Reaction Buffer. The colored reaction buffer contains red and orange dyes, which separate during electrophoresis and provide quick reference points for monitoring the mobility of the DNA samples in the gel. The colored reaction buffer can be loaded directly onto an agarose gel for analysis without the need for separate gel-loading buffer. The presence of the dyes has no effect on routine enzymatic manipulations, although extremely rare exceptions may exist.
Since the colorless reaction buffer does not contain reference dyes, it is suitable for use when reaction products will be used directly for down-stream processes involving absorbance or fluorescent detection. The specificity and performance of MangoTaq DNA Polymerase can be further improved with the use of 3% DMSO, which is designed for GC or AT-rich DNA, "dirty" templates or sequences with a high level of secondary structure.
Applications
High throughput applications
Suited to a wide range of PCR assays
Products suitable for TA cloning
Direct loading
Fig. 1 Amplification of a 1 kb fragment from l DNA using MangoTaq DNA Polymerase and supplier Q Taq DNA Polymerases.
A 1 kb l DNA fragment was amplified using MangoTaq and Taq from supplier Q. The DNA fragment was amplified from a 5-fold serial dilution of l DNA with an initial concentration of 0.5 ng (lane 1), 0.1 ng l DNA (lane 2), 0.02 ng lDNA (lane 3), 4 pg l DNA (lane 4). PCR was performed in 50 µL reaction mixtures containing 1.5 mM MgCl2. HyperLadder 1kb (M).
Fig. 2 Amplification of a range of fragments from different human genes using MangoTaq DNA Polymerase and supplier Q Taq DNA Polymerase.
The amplification products are as follows: 119 bp (43% GC) from human glucocerebrosidase gene (1), 321 bp (37% GC) from angiotensin receptor II gene (2), 635 bp (56% GC) from rhodopsin gene (3), 762 bp (33% GC) from b-globin gene (4), 1200 bp (54% GC) from a-1-antitrypsin gene (5). PCR was performed in 50 µL reaction mixtures containing 50 ng human genomic DNA and 1.5 mM MgCl2. HyperLadder 50bp (M).
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 of the reagents is retained until the expiry date indicated on the outer box label.
Shipping conditions
On Dry Ice or Blue Ice.
One unit will incorporate 10 nmoles of dNTPs in 30 min at 72°C
""The best performance was exhibited by the MangoTaq DNA polymerase, ...
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University of Hohenheim, Stuttgart, Germany
"The best performance was exhibited by the MangoTaq DNA polymerase, which was the only polymerase which was able to amplify the 620bp amplification product from the 102 year old sample."
No, the dyes and composition of MangoTaq Reaction Buffer 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 two colored dyes present in this buffer are completely inert and thus will have no effect on downstream applications. An exception is the quantification of PCR products in colored buffers with photometric methods or fluorescence assays. We cannot exclude the impact of these dyes on quantification results and suggest the purification of these samples. If you are concerned about these dyes interfering in your specific applications, we would recommend the cleanup of your samples using the ISOLATE II PCR & Gel Kit or SureClean Plus.
At Meridian 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.
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.
All Meridian 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.
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.
