TaqRobat Native Taq MantisGreen 2X Master Mix is an optimized, ready-to-use mix that contains dNTPs, buffers, stabilizers, MantisGreen loading dyes and our TaqRobat recombinant 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. TaqRobat 2X Master Mix provides robust amplification over a wide range of templates up to 5kb. 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	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.