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Thermo Scientific™ HotStart-IT Binding Protein
Description
HotStart-IT Binding Protein is the active component in the novel hot start technology known as primer sequestration. In primer sequestration, hot start PCR methods reduce or eliminate non-specific primer-extension products formed at lower temperatures during assembly of PCR reactions. At these less stringent annealing temperatures, primers may bind non-specifically, which often leads to unwanted amplification products and primer-dimers. To resolve this problem, we have produced a high-quality DNA binding protein that is especially useful at sequestering primers at lower temperatures, making them unavailable for use by a polymerase (see figure).
Features
- Room temperature reaction setup
- Minimizes amplification of non-specific products and primer-dimers (see figure)
- Ideal for complex templates and multiplex reactions
- No extensive heating step required
- Free from detectable non-specific nucleases
- Technology is portable to a polymerase of choice
This primer-sequestration technique effectively blocks DNA synthesis from mis-priming events at lower temperatures. Following the initial denaturation step during PCR, the protein is inactivated, and the primers are free to participate in the amplification reaction.
HotStart-IT Binding Protein performs well in many standard PCR reaction buffers and has been designed for PCR applications that demand high specificity and sensitivity. PCR with Taq DNA Polymerase and HotStart-IT Binding Protein shifts production of primer-dimers to a specific target of 306 bp from 1 ng of human genomic DNA relative to Taq DNA Polymerase alone.
Application
- Hot-Start PCR amplification
Notes
- One microgram of HotStart-IT Binding Protein sequesters about 5 pmol of primers.
- HotStart-IT Binding Protein is sourced from recombinant protein expressed in E. coli.
Specifications
Specifications
| For Use With (Application) | Hot-start PCR |
| Content And Storage | • HotStart-IT Binding Protein, 400 μg Store at -20°C. |
| Shipping Condition | Dry Ice |
| Compatible Buffer | 1X PCR Reaction Buffer |
| Quantity | 400 μg (200 x 25 μL rxns) |
| Product Type | HotStart-IT Binding Protein |
For Research Use Only. Not for use in diagnostic procedures.
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