rTaq Plus 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 DNA Polymerase is contained in 4 ㎕ of 5x Master Mix.rTaq Plus DNA Polymerase is a thermostable DNA polymerase that designed for a reliable amplification of long and complex targets with a robust yield in an extremely short extension time. Due to its high elongation rate (about 100 bp/sec) and high processivity (about 250 bases), long sized products can be generated within relatively shorter running time compared to any other commercially available enzymes (5 kbp within 60 sec extension).Recombinant rTaq Plus DNA Polymerase is purified from a modified recombinant E.coli strain. The enzyme catalyzes the incorporation of nucleotides into duplex DNA in the 5' to 3´ direction in the presence of Mg²⁺ at 70-80°C. rTaq Plus DNA Polymerase has no detectable exonuclease activity (5' to 3´ or 3' to 5´) and generates a 3´dA overhang, suitable for a TA cloning of PCR products.rTaq Plus DNA Polymerase is free from any nucleic acid which can be contaminated from expression host during purification process. That provides robust conditions for amplification of gram-negative bacterial genes, and profiling of these.
QC tests- Performance tests, genomic DNA contamination test, confirmation test for the absence of endo and exonucleases. Storage Condition- Store at -20°C.- rTaq Plus 5x PCR Master Mix is stable for at least 2 years at recommended storage condition. Standard Protocol1.Prepare 20 ㎕ PCR solution as follows:PCR grade distilled water : - ㎕rTaq Plus 5x PCR Master Mix : 4 ㎕Primer (10 pmol/㎕) : 0.5 ㎕ eachTemplate DNA : 0.1-10 ngAdjust final vol. to 20 ㎕ with PCR grade distilled water2. Set PCR cycling as follows :Initial denature at 95°C : 3 minDenature : 95°C, 10 secAnneal : Tm-4°C, 10 secExtend : 72°C, 10 sec/kbp* Set 25-35 PCR cycles for effective amplification* You can also use two step cycle 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.