2x Taq PCR Mix, Biomiga

2x PCR Mix is a ready-to-use and ready-to-load reaction cocktail for PCR amplification up to 4 kb. It’s a 2X concentrated formulation that contains all necessary components including Magic Taq DNA polymerase, magnesium, dNTPs, glycerol, and blue color tracking dye, except primers and template. Reagents sufficient for 100 or 500 or 5000 PCR amplification reactions of 50 µl reaction volume.

 

Features

• High yield, high sensitivity

• Superior reliability and robustness

• Convenient and time saving: ready-to-use and ready-to-load PCR Mix

 

Product Qualification

2x PCR Mix is functionally tested for amplification of a 2-kb and 4-kb fragment from a single copy gene in human genomic DNA.

 

Recommended PCR Reaction Protocol

The following protocol is suggested as a starting point.

Components

25 ul Rxn

50 ul Rxn

Final Concentration

PCR Mix (2X)

12.5 ul

25 ul

1X

Forward Primer (10 uM)

0.5 ul

1.0 ul

200 nM (Variable:100-500 nM)

Reverse Primer (10 uM)

0.5 ul

1.0 ul

200 nM (Variable:100-500 nM)

Template DNA

x ul

x ul

Variable (fg - ug)

Final Volume (ul)

25 ul

50 ul

 

 

1. Assemble the reaction on ice.

2. Cap reaction vessels and load in thermal cycler at 94°C.

3. Incubate tubes in a thermal cycler at 94°C for 30 s to 1 min to completely denature the template.

4. Perform 25-40 cycles of PCR amplification as follows:

▪ Denature 94°C for 15–30 s

▪ Anneal 55-60°C for 15–30 s

▪ Extend 72°C for 1 min per kb

▪ Hold at 4°C until use

5. Analyze PCR products by gel electrophoresis

 

Store at -20°C

2x PCR Mix is stable for 1 year when stored at -20°C. It may be stored at 4°C to avoid the necessity of repeated thawing the mix before assembling the PCR. No detectable reduction of PCR performance was observed after storage for 12 months at 4°C. Repeated freeze-thaw cycles do not impair PCR performance.