TaqTivate Hot Start Taq Master Mix is an optimized, ready-to-use mix that contains dNTPs, MgCl2, optimized buffers and our TaqTivate recombinant hot start Taq DNA polymerase. It is supplied in a 2X formulation that requires only the addition of primers, template and water. TaqTivate requires a 95C activation time of only two minutes for faster cycling reactions. Chemical modification keeps TaqTivate free of contaminating DNA from biological modifying agents such as antibodies. TaqTivate 2X Master Mix provides robust amplification of templates up to 5kb and reduces non-specific DNA amplification due to primer-dimer formation. Suitable for Standard Endpoint PCR, Colony PCR, Low Copy PCR, Genotyping, and TA Cloning. Also available containing MantisGreen dual-color loading dyes.
Reaction setup: The following setup is recommended for a 50µl reaction but can vary depending on the template and primers being used.
Component | Volume | Final Concentration |
Master Mix | 25µl | 1X |
5' Primer, 10µM | 0.5-5µl | 0.1-1µM |
3' Primer, 10µM | 0.5-5µl | 0.1-1µM |
DNA Template | 0.1-5µl | >1ng |
Nuclease Free Water | QS to 50µl |
Thermal cycling conditions: The following general cycling conditions are recommended but can vary depending on the template and primers being used.
Cycling Step | Temperature | Holding Time | Cycles |
Initial Denaturation and Hot Start Activation | 94°C | 30sec-2min | 1 |
Denaturation | 94-96°C | 15-30sec | 20-30 |
Annealing* | 55-65°C | 15-60sec |
Extension | 70-72°C | 1min per kb |
Final Extension | 70-72°C | 0-10min | 1 |
*Annealing temperature will depend on primer length and composition. Generally, begin 5°C below primer Tm.