Description
Purity:>95%, by SDS-PAGE visualized with Coomassie® Blue Staining.Taq DNA Polymerase is a thermostable enzyme that synthesizes DNA from single-stranded templates in the presence of dNTPs and a primer. The enzyme consists of a single polypeptide with a molecular weight of 94 kDa. It has a 5´→3´ DNA polymerase activity and a 5´→3´ exonuclease activity.Source：Recombinant enzyme is purified from the cloned Thermus aquaticus DNA polymerase gene expressed in E. coli.Unit definitionOne unit of Taq DNA Polymerase is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into DNA in 30 min at 74°C.Product content：1. Taq DNA Polymerase ( 5 U/µL )2. 10×Taq Buffer（Mg2+Plus）：200mM Tris-HCl,500mM KCl,20mM MgCl2,1mM DTT,pH9.3PCR Reaction Setup：Component50-µL rxn5×Taq Buffer(Mg2+ Plus)10µLdNTP Mixture(2.5mM each)4µLTemplate DNAPrimer F0.2-1.0 µMPrimer R0.2-1.0 µMTaq DNA Polymerase(5U/µL)0.2µLddH2OUp to 50µLIncubate reactions in a thermal cycler：StepTemperature（℃）TimeCyclesInitial Denaturation955min1Denature9530s25-35Anneal~55(depending on primer Tm)30sExtend721min/kbFinal Extension7210min1Hold4indefinitely1Optimization Strategies：Different PCR reaction conditions, including temperature, time and number of cycles, should be set according to different template, primer, length of PCR product and GC content.The time setting of STEP4(extension) should be set according to the length of PCR products, usually the extension time of each kb product is 1min.For initial PCR, the number of cycles can be set to 35 to ensure that the expected PCR product can be amplified as much as possible. The number of PCR reaction cycles that need to be semi-quantitative or quantitative must be properly optimized so that the PCR reaction does not reach a plateau