BIOTAQ^™ DNA Polymerase

Please see the following table for the appropriate error rates: BIOTAQ, IMMOLASE, MangoTaq, MyTaq DNA Polymerase 1.1 x 10-4 base substitutions/bp (Tindall and Kunkel, 1988) 2.4 x 10-5 frameshift mutations/bp (Tindall and Kunkel, 1988) 2.1 x 10-4 errors/bp (Keohavang and Thilly, 1989) 7.2 x 10-5 errors/bp (Ling et al., 1991) 8.9 x 10-5 errors/bp (Cariello et al., 1991) 2.0 x 10-5 errors/bp (Lundberg et al., 1991) 1.1 x 10-4 errors/bp (Barnes, 1992) ACCUZYME 1.6 x 10-6 errors/base (Lundberg et al., 1991). VELOCITY 4.4 x 10-7 errors/base (Frey & Suppmann, 1995).

BIOTAQ DNA Polymerase is a fast enzyme, which will extend fragments at 15-30 s/Kb, dependent on template amplified.

Yes, BIOTAQ 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.

BIOTAQ will comfortably amplify fragments of up to 5 kb with genomic templates.

BIOTAQ DNA Polymerase will extend between 50-80°C, however optimal extension will occur at 72°C.

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.

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.