| 摘要 |
PURPOSE: A hot start PCR(polymerase chain reaction) compound containing pyrophosphate and pyrophosphatase is provided, which inhibits production of non-specific amplified products, so specific amplified products can be produced effectively, and a PCR method to inhibit non-specific amplification and production of primer-dimer with the same is also provided. CONSTITUTION: A process for the preparation of hot start PCR compound containing pyrophosphate and pyrophosphatase comprises of: measuring a concentration of pyrophosphate inhibiting PCR; measuring an activity unit of pyrophosphatase emitting Mg2+ ion; adding pyrophosphate(0.3-5mM, desirably 0.5mM) and pyrophosphatase(0.0001-0.01U, desirably 0.0003U per PCR compound 20microliter) to PCR compound containing tris-HCl(pH 8.3), KCl, MgCl2, DTT, BSA, 4 kind of dNTP and DNA polymerase, and mixing(colors-bromophenol blue, xylene cyanole, bromocresol red, or cresol red- and stabilizer-gelatin, bovine serum albumin, The sit, PEG-8000, or polyol- can be added); confirming inhibition of production of non-specific amplified products by hot start PCR(polymerase chain reaction) compound containing pyrophosphate and pyrophosphatase.
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