Request a sample of IsoFast® Hot Start Bst Colour Mix or Polymerase

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IsoFast® Hot Start Bst Colour Mix is designed as a cutting-edge solution in molecular biology, specifically developed for field-based DNA detection and amplification. This innovative product stands out in the field of isothermal amplification, offering rapid, sensitive, and specific DNA detection, ideal for various applications, including point-of-care microbial DNA testing.

IsoFast® Hot Start Bst Colour Mix is powered by Bst DNA polymerase combined with AptaLock™  hot start technology and a pH-sensitive dye. This mix enables isothermal DNA amplification with direct colourimetric positive/negative screening.

Proprietary hot start reduces non-specific amplification, increases specificity, improves sensitivity and enables completion of isothermal amplification runs in as little as 30 minutes. IsoFast® Hot Start Bst Colour Mix, is a one tube formulation of Bst polymerase with all buffers and dNTPs for reduced pipetting, including a colour indicator for colourimetric readout. The mix is also available as a separate Bst polymerase and a colour buffer system, allowing you to tailor assay conditions in combination with a colourimetric readout. This is ideal for novel detection methods and synthetic biology detection schemes.

Complete the form to test IsoFast® Hot Start Bst Colour Mix or Colour Polymerase in your workflow.

IsoFast Hot Start Colour Mix Plate

IsoFast® Hot Start Bst Colour mix offers a rapid and reliable colourimetric readout at cold and ambient reaction setup. N.b. The image shown does not represent actual experimental results.

What is IsoFast®?

IsoFast® Bst Polymerase is a recombinant protein expressed in E. coli and represents the large fragment of Geobacillus stearothermophilus (formerly known as Bacillus stearothermophilus) DNA Polymerase. This portion of the protein catalyses the 5′-3′ synthesis of DNA and has strand displacement activity but does not contain the 5’-3’ exonuclease domain.1

How does isothermal amplification with IsoFast® work?

Strand displacement refers to the ability of an enzyme to dissociate the hydrogen bond of double stranded template DNA encountered downstream, essentially unzipping the DNA as the complementary strand is synthesised. IsoFast® Bst Polymerase displays strong strand displacement activity and is suitable for amplification methods including whole genome amplification, multiple displacement amplification and isothermal amplification. DNA synthesis is performed at a constant temperature, and we recommend running the reaction at 65 ºC. However IsoFast® Bst Polymerase works well over a broad temperature range (55 ºC to 70 ºC, see Figure 4), giving a wider range of reaction conditions in order to optimise primer annealing and strand displacement. The enzyme is heat inactivated at 80 ºC.

Preventing non-specific amplification in 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 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:

Hot start techniques can also improve the sensitivity of isothermal amplification assays. By minimizing 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.

Key Features:

  • Fast Colour Readout: The mix provides a quick colour readout for positive/negative DNA testing, crucial for rapid diagnostics.
  • AptaLock™ Hot Start: This feature ensures ultra-sensitive DNA detection, minimizing non-specific amplification and false positives.
  • Rapid Polymerisation: It offers faster results, with the potential to detect as little as 3 target copies per μL in as little as 10 minutes.
  • Temperature Versatility: High activity across a broad temperature range (55-70 °C), suitable for diverse experimental conditions.
  • Isothermal Amplification: PCR-free, direct, colour-based readout of DNA targets, making it highly efficient and user-friendly.

Why Choose IsoFast® Hot Start Bst Colour Mix for microbiology applications?

  • Enhanced Sensitivity and Speed: The mix is designed for early target detection, improving the speed and sensitivity of DNA amplification.
  • Flexibility: Available as a preassembled mix for ease of use or as a separate enzyme and buffer system for full assay customization, use in a heat block or thermocycler.
  • Hot Start Technology: Critical in isothermal amplification, it prevents non-specific amplification and enhances the assay’s specificity and sensitivity.

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