TaqTivate Hot Start Taq MantisGreen Master Mix is an optimized, ready-to-use mix that contains dNTPs, MgCl2, optimized buffers, MantisGreen loading dyes and our TaqTivate recombinant hot start Taq DNA polymerase. Reactions can go directly from PCR tube to gel. The mix is a 2X formulation that requires only the addition of primers, template and water. MantisGreen dual-color loading dyes do not inhibit PCR and make monitoring progress easy during electrophoresis. The dyes run outside the range of typical PCR products and therefore do not obscure visualization. The blue dye runs at 4kb and the yellow dye runs under 25bp. 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 direct-to-gel Standard Endpoint PCR, Colony PCR, Low Copy PCR, Genotyping, and TA Cloning. Also available without 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.