illustra™ Hot Start Mix, Ready-To-Go™ is a pre-formulated and pre-dispensed, freeze-dried PCR reagent mix, including high quality PuReTaq™ DNA polymerase and a hot start activator protein, for increased specificity and reproducibility of PCR amplifications. The Ready-To-Go™ beads are provided in either 0,5 or 0,2 ml tubes that are compatible with most thermal cyclers.
- Recombinant hot start activator protein sequesters primers prior to PCR, effectively reducing non specific priming and primer-dimer formation
- Avoids the risk of contamination from a mammalian source associated with hot start antibodies
- Quick hot start activation preserves sample integrity by avoiding extensive pre-cycling heat treatment
- Long term ambient-temperature-stable: No freezer space required; less energy consumption for shipping and storage
- Validated for real time PCR
Kit includes room-temperature stable beads containing 1.75 units of recombinant PuReTaq DNA polymerase, dATP, dCTP, dGTP, dTTP, stabilizers, BSA and reaction buffer. When a bead is reconstituted to a 25 μl final volume, the concentration of each dNTP is 200 μM and the concetration of MgCl₂ is 1.5 mM.
The additional reagents required are water, primers, and template DNA. MgCl₂ can be added, allowing users to customize this reagent to their specific needs. The beads are provided predispensed into either 0.2 or 0.5ml PCR tubes compatible with most thermal cyclers. The 0.2 ml tubes come assembed in a 96-well (8×12) plate format that allows individual strips of eight tubes to be easily removed. This flexibility allows use of the entire 96-well plate, strips of eight, or individual 0.2 ml tubes. Avoids the risk of contamination from a mammalian source associated with hot start antibodies.
Quick hot start activation to preserve sample integrity by avoiding extensive precycling heat treatment.
Long-term ambient-temperature stable: No freezer space required - Less energy consumption for shipping and storage.
This Ready-To-Go format provides reproducible and reliable performance for demanding PCR applications in which high specificity and sensitivity are essential to success.
The system is based on a novel hot start method that uses a hot start activator protein to sequester primers prior to PCR, thereby making them unavailable for nonspecific priming during the reaction preparation. This method requires only a quick 2 min initial PCR activation step. In contrast, other available hot start PCR kits that are based on chemical inactivation require prolonged incubation at high temperatures, which may damage the DNA template by heat-induced depurination. The net result from using Hot Start Mix RTG is a more robust and reliable amplification of the target amplicons with increased yield when compared with conventional hot start master mix products. Furthermore, the mix does not contain Taq antibody and this eliminates the risk of mammalian-source contamination.