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.