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 hot start PCR
• High-yield PCR
• Fast PCR
• Multiplex PCR
• qPCR and RT-qPCR

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 HS DNA Polymerase works very well with difficult samples like GC rich or bisulfite converted DNA. The hot start PCR is especially 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.