Hot start Taq DNA Polymerase for qPCR is designed for Real-Time PCR and Hot-start PCR. A
special inhibition the reaction at room temperature until after the first denaturation step. This prevents primerdimers
and other artefacts.
PowerAmp 2X PCRmix-Green is optimized mixture contain of Hot Start Taq enzyme, reaction buffer, dNTP,
enhancer as 2-fold concentration. PowerAmp 2X PCRmix-Green is designed to allow the user for quick ,
easy preparation of reaction mixture.The PowerAmp 2X PCRmix-Green can be amplification PCR products
up to 5 kb and the products can be directly cloning into T-vector.
Primers: Use 0.3 μM per primer as a general starting point. For larger amounts of template (e.g., 200 ng genomic
DNA), increasing the concentration up to 0.5 μM per primer may improve yield.
Annealing Temperature: The annealing temperature is slightly higher than with typical PCR. The optimal annealing
temperature should be ~2°C lower than the Tm of the primers used. A range of 58–68°C is recommended.
Extension Time: As little as 30 seconds per kb is suitable for most targets. Use up to 60 seconds per kb for
1.Thaw the 2x Hot start mix at room temperature. Vortex
the 2x Hot start mix and then spin it briefly in a micro
centrifuge to collect the material in the bottom of the tube.
2. Prepare one of the following reaction mixes on ice:
|2x Hotstart mix||12.5 ul|
|Primer1 (20 pmol)||1-2 ul|
|Primer2 (20 pmol)||1-2 ul|
|ddH2O||Up to 25 ul|
3. If necessary you can scale up your volume
Program the thermal cycler as follows
After cycling, maintain the reaction at 4°C. Samples can
be stored at –20°C until use.
Analyze products using standard agarose gel electrophoresis.