Standard thermostable polymerases (e.g. Taq) have optimal performance around 70 °C. Yet, moderate enzymatic activity at room temperature creates unspecific products.
Hot Start Technology increases specificity and lets you set up reactions at room temperature. Inhibition of Hot Start Polymerases at ambient temperature prevents unspecific reactions before heating begins. Heat-induced inhibitor release takes place during the first step of PCR (Fig. 1).
Hot Start Polymerases reduce the generation of undesired fragments and allow for setup at room temperature. Chemically modified polymerases need a heat activation step which can lead to DNA damage. In contrast, aptamers or antibodies inhibit Jena Bioscience Hot Start Polymerases and activation occurs during the initial denaturation step of the PCR, eliminating DNA damage.
Hot Start Polymerase Ab+ is most strongly inhibited at room temperature and gives maximum specificity. However, antibodies may be from animal origin. In order to prevent mammalian nucleic acid contamination, Jena Bioscience offers Hot Start Polymerase, aptamer blocked based on a fully synthetic inhibitor.
| Antibody based Hot Start Technology | Aptamer based Hot Start Technology |
|---|---|
| Hot Start Polymerase Ab+ (PCR-213) Thermus aquaticus, recombinant, E. coli |
Hot Start Polymerase Apta+ (PCR-212) Thermus aquaticus, recombinant, E. coli |
| Hot Start Core Kit Ab+ (PCR-216) Kit of Hot Start Pol, dNTPs and reaction buffer |
Hot Start Core Kit Apta+ (PCR-215) Kit of Hot Start Pol, dNTPs and reaction buffer |
| Crystal Hot Start Master (2x) (PCR-167) Master mix, ready-to-use |
|
| Ruby Hot Start Master (2x) (PCR-165) Master mix, ready-to-use for direct-to-gel analysis |
E-mail Bürk for service inquiries or further information at pcr@jenabioscience.com !
