IsoFast™ Hot Start Bst Colour Mix – Witec AG

Applications

• Colourimetric Isothermal Amplification
• Colourimetric LAMP
• Positive/negative DNA testing
• Rapid target screening
• Point-of-care testing

Hot start isothermal amplification

Hot start is important in isothermal amplification techniques because it helps to prevent non-specific amplification and false-positive results. Isothermal amplification methods, such as loop-mediated isothermal amplification (LAMP), rely on specific primers and enzymes to amplify a target sequence at a constant temperature, typically between 60°C and 65°C. These techniques are widely used in various applications, including molecular diagnostics and pathogen detection.

Why is hot start crucial in isothermal amplification?

• Preventing non-specific amplification in Bst polymerase isothermal reactions: In isothermal amplification, it’s essential to ensure that the amplification process only targets the specific sequence of interest. Non-specific amplification can occur when primers bind to unintended regions of the template or when amplification enzymes prematurely initiate amplification. Hot start techniques involve inhibiting the amplification reaction until the reaction reaches the desired temperature. This prevents the formation of non-specific products during the initial stages of the reaction when the temperature is still below the optimal amplification temperature.
• Enhancing Bst polymerase isothermal amplification specificity: By preventing premature amplification, hot start methods increase the specificity of the isothermal amplification reaction. This means that the reaction is less likely to amplify non-target sequences, reducing the risk of false-positive results.
• Improving isothermal amplification sensitivity with Bst polymerase: Hot start techniques can also improve the sensitivity of isothermal amplification assays. By minimising non-specific amplification, more of the available reagents are reserved for amplifying the target sequence, increasing the efficiency of the reaction and the detection limit.