rTaq Plus HOT 5x PCR Master Mix is a ready-to-use premix containing all the components essential for a PCR and agarose gel electrophoresis (DNA polymerase, dNTP, reaction buffer, glycerol, bromophenol blue, and stabilizer). PCR can be performed simply by adding primer pair and template.
As Master Mix is supplied as a 5x concentration format, users should adjust a final reaction to 1x (if final reaction volume is 20 ㎕, 4 ㎕ of 5x Master Mix should be added).
One unit of rTaq Plus HOT DNA polymerase is contained in 4 ㎕ of 5x PCR Master Mix.
rTaq Plus HOT DNA Polymerase in Master Mix is suitable for a high fidelity amplification of DNA fragments.
This enzyme is designed for a reliable amplification of long (up to 10 kbp for lambda DNA) and complex targets with a robust yield and high specificity.
- Performance tests, genomic DNA contamination test, confirmation test for the absence of endo and exonucleases.
- Store at -20°C. HiPi Plus 5x PCR Master Mix is stable for at least 2 years at recommended storage condition.
1. Prepare 20 ㎕ PCR solution as follows:
PCR grade distilled water : - ㎕
rTaq Plus HOT 5x PCR Master Mix : 4 ㎕
Primer (10 pmol/㎕) : 0.5 ㎕ each
Template : 0.1-10 ng
Adjust final vol. to 20 ㎕with PCR grade distilled water
2. Set PCR cycling as follows :
Initial denature at 95°C : 3 min
Denature 95°C 30 sec
Anneal Tm-4°C 30 sec
Extend 72°C 30-60 sec/kbp
* Set 25-40 PCR cycles for effective amplification
* You can also use two step cycle for > 5 kbp amplification (denaturation at 95°C and annealing/extension at 68°C)
3. You can analyze PCR products by direct loading into agarose gel because PCR Master Mix
contains glycerol and bromophenol blue (blue color) essential for a gel loading.