PCR Biosystems IsoFast Hot Start isothermal mixes

PCR Biosystems has launched a range of Bst polymerase reagents that offer high sensitivity, speed and specificity in all isothermal amplification applications. IsoFast Hot Start Bst Polymerase and associated ready mixes use proprietary AptaLock hot start technology to minimise non-specific amplification, leading to specific and sensitive results in as little as 30 min. A colorimetric mix also enables analysts to detect results visually, reducing equipment requirements. With these technologies, labs can now get results more quickly, unlocking high-throughput testing.

A common issue with isothermal mixes is non-specific amplification — where primers bind to unintended DNA targets — because amplification often takes place at a fixed temperature which may favour non-specific primer interactions, contrary to PCR. Isothermal mixes can also lead to primer dimer formation, where primers associate with each other and are extended by the polymerase. To avoid these issues, users must design primers carefully, and evaluate many more primer pairs for a single target than they would for standard PCR.

To help alleviate these concerns, PCR Biosystems’ IsoFast reagents use AptaLock, a reversible hot start technology that inhibits polymerase activity below 40°C. As such, both non-specific amplification and primer dimer formation are minimised. Once the mixture is activated on heating, results can be obtained in as little as half an hour, allowing analysts to achieve sensitive, specific and rapid detection of DNA targets. High-throughput testing is also possible, as reaction plates can be set up at room temperature without any issues.

The colour mix contains a pH-based dye that changes colour when the desired target is amplified. As a result, the mix enables labs to visually identify positive or negative results without the need for plate readers or fluorometric PCR instruments. The mix is therefore beneficial for regions where point-of-care or field-testing laboratories lack adequate facilities and instrumentation.