Standard thermostable polymerases (e.g. Taq) show an optimal performance around 70°C. Yet, moderate enzymatic activity at room temperature can create unspecific by-products.
Hot start technology significantly reduces the risk of generating such unwanted fragments by inhibiting the polymerases at ambient temperature and activating them during the first PCR cycle. This prevents unspecific reactions before heating begins and allows for a PCR setup at room temperature (Fig. 1).
Fig. 1: Detection of genetically modified Roundup Ready® soy (Monsanto) down to 0.1 % in soybean flour was compared for Hot Start Pol and conventional Taq Pol. Formation of by-product and associated risk of false positive detection is greatly reduced by Hot Start Pol. Assay: Amplification of genomic DNA, 210 bp fragment of soy transgene, 150 ng DNA / reaction, 1.25 units Hot Start Pol / reaction, 95°C, 2 min; 35x (95 °C, 30 sec; 56 °C, 30 sec; 72 °C, 30 sec); 72 °C, 4 min.
Jena Bioscience offers you two options for your Hot Start Polymerase: either inhibited by an apatamer (Hot Start Polymerase Apta+) or by an antibody (Hot Start Polymerase Ab+). As antibodies are of animal origin and bring a minimal risk of mammalian nucleic acid contamination, Jena Bioscience introduced the Hot Start Polymerase Apta+ for you - an alternative based on a fully synthetic inhibitor.
Both products are high quality Taq Polymerases that are most strongly inhibited at room temperature while giving maximum specificity. They can be easily activated during the initial denaturation step of the PCR, therefore avoiding the risk of DNA damage due to prolonged heat treatment.
Also available as Master Mixes or Core Kits (Hot Start Polymerases).
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