HiPi Super 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 HiPi Super DNA Polymerase is contained in 4 ㎕ of 5x Master Mix.Thermostable HiPi Super DNA Polymerase is suitable for an amplification of < 40 kb DNA fragments with a high fidelity and specificity.It is highly effective in long PCR and multiplex PCR.HiPi Super DNA Polymerase catalyzes the incorporation of nucleotides into duplex DNA in the 5´->3´ direction in the presence of Mg²⁺ at 70-80°C. HiPi Super DNA Polymerase is a mix of DNA polymerase showing 3´->5´ exonuclease (proof-reading) activity and Taq DNA Polymerase. Due to its high elongation rate, long sized products can be generated within relatively shorter running time compared to any other commercially available enzymes (10 kbp within 30 sec extension).This enzyme is designed for a reliable amplification of long (up to 40 kbp for lambda DNA and 20 kbp for human genomic DNA) and complex targets with a robust yield and a high accuracy.HiPi Super DNA Polymerase generates a mixture of PCR products with blunt end and 3´dA overhangs.
- Performance tests, genomic DNA contamination test, confirmation test for the absence of endo and exonucleases.
Storage Condition- Store at -20°C. HiPi Super 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 : - ㎕HiPi Super 5x PCR Master Mix : 4 ㎕Primer (10 pmol/㎕) : 0.5 ㎕ eachTemplate : 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 5 secAnneal : Tm-4°C 5 secExtend : 72°C 5 sec/1kbp* Set 25-35 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.