Taq DNA Polymerase
Taq DNA Polymerase is a combination of next-generation DNA polymerase and a novel buffer system that has been designed to deliver very high yield, fast PCR amplification over a wide range of PCR templates. It has been developed to give very robust amplification, allowing it to perform well with most PCR testing, including challenging templates and in the presence of PCR inhibitors.
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Robust PCR amplification of GC-rich human genomic DNA
A serial dilution in duplicate of human genomic DNA (1 μg, 200 ng, 100 ng, 50 ng, 25 ng and 12.5 ng, 1-6 respectively) was used with primers and Taq DNA Polymerase to amplify a 450 bp fragment of the human myc gene. The section of gene used has 61% GC content and was selected to demonstrate the high yield and sensitivity of the Taq DNA Polymerase, making it the perfect choice for the amplification of complex PCR templates.
Taq DNA Polymerase, MDX001
Available in 0.5 mL (500 Units) or 10 mL (50,000 Units) aliquots
Taq DNA Polymerase is provided with an optimized buffer system containing dNTP, MgCl2, stabilizers, and PCR enhancers at optimal concentrations which eliminates the need for optimization. The DNA polymerase has been developed to give more robust amplification than other polymerases allowing it to perform well with challenging templates in the presence of PCR inhibitors, making it ideal for fast PCR reactions across a range of templates, without compromising specificity or yield.
|Description||Very high-performance polymerase, supplied with an optimized, novel buffer system, containing dNTPs, MgCl2 and enhancers, to give superior amplification.|
|Appearance||Clear, colorless solution|
|Sample type||cDNA, DNA|
|Mix stability||See outer label|
|Functionality||PCR with a dilution series of human genomic DNA, single, distinct band was observed|
|DNA Contamination||None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets.|
|DNase Contamination||No detectable degradation|
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FAQs: Taq HS DNA Polymerase
Decreasing the template concentration will help to minimize the concentration of inhibitors, the amount of DNA polymerase and primer can also be increased. In some cases you may also have to add a pre-treatment step if the DNA is not accessible.
These terms refer to parameters to be considered when performing PCR testing.
Yield: The amount of DNA produced in a PCR reaction. High yield increases sensitivity.
Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand. For standard PCR this is not important, but for sequencing or expression a high-fidelity mix (such as High-Specificity Pfu HS Mix (MDX006)) may be required.
Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified (the amplicon). Taq HS DNA Polymerase will work with amplicons up to 5 kb.
Specificity: A measure of the unwanted by-products generated in a reaction. This is the reason for having hot start PCR, this prevents miss-priming at low temperatures and non-specific amplification
Taq DNA Polymerase has been validated with a broad range of PCR templates including DNA extracted from human, animal and plant samples, making it the ideal choice for the following applications:
• End-point PCR testing
• High-yield PCR testing
• Fast PCR testing
• Multiplex PCR testing
• qPCR and RT-qPCR testing
Yes, Taq DNA Polymerase can be used for quantitative PCR amplification (qPCR), but it will require a hot-start antibody (MDX014), qPCR buffer such as Fast qPCR Buffer, 4x (MDX033), or for RT-qPCR, a one-step buffer such as 1-Step qPCR Buffer, 4x (MDX034) and reverse transcriptase such as MMLV-RT (MDX044).
This depends on the length of the amplicon and the complexity of the template. For fast PCR, the initial denaturation will remain 1 minute, but with low complexity template such as plasmid DNA, an annealing of 5 seconds and extension time of 10 seconds is sufficient for amplicons up to 1 kb. In order to find the fastest optimal condition, for more complex or longer amplicons, we suggest incrementally increasing the extension time successively up to 30 s/kb.
Yes, Taq DNA Polymerase works very well with difficult samples like GC rich or bisulfite converted DNA. However a hot-start to the PCR (such as MDX014) is important, as it decreases primer dimer and unspecific amplification.
If optimization is required, we would suggest starting with the annealing temperature and with bisulfite conversion, the process can fragment the DNA, so it may be useful to increase the amount of template and polymerase.
The enzyme has an error rate of approximately 1 error per 2.0 x 10⁵ nucleotides incorporated. If you are looking for a DNA polymerase with a lower error rate, for next-generation sequencing (NGS) library or expression a high-fidelity DNA polymerase (such as High-Fidelity Pfu (MDX003)) or mix (such as High-Specificity Pfu HS Mix (MDX006)) should be used.
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