IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes IsoFast® Hot Start Bst Polymerase & Mixes

IsoFast® Hot Start Bst Polymerase & Mixes

Bst DNA Polymerase combined with reversible hot start technology AptaLock™ powers our IsoFast® Hot Start Bst reagents. These products are ideal for all homogeneous amplification processes, providing more sensitive and reliable results faster than ever before. IsoFast® Hot Start Bst Polymerase is a recombinant version of the large fragment of Geobacillus stearothermophilus (formerly known as Bacillus stearothermophilus, Bst) DNA Polymerase. It is designed to maintain 5'-3' polymerase activity but lacks 5'-3' exonuclease activity. The strong strand displacement capability of the enzyme eliminates the need for the denaturation process required by standard PCR. Combined with powerful hot-start technology, designed to minimize the formation of primer dimers and non-specific target amplification, this new formulation is ideal for non-PCR-based amplification. Characteristics: AptaLock™ hot start for ultra-sensitive target DNA detection Rapid polymerization for faster results (just about 10 minutes) Detection down to 3 target copies per μL Ideal for both cold and room temperature setups Enhances speed and sensitivity for early target detection High activity over a wide temperature range from 55-70 Available with and without fluorescent dye, and in a pre-mixed 2x format Available formats for colorimetric assays.
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CAT CodeProductSizePrice
PB80.40-01IsoFast® Hot Start Bst Polymerase1600 UnitsContact us
PB80.40-08IsoFast® Hot Start Bst Polymerase8000 UnitsContact us
PB80.41-01IsoFast® Hot Start Bst Polymerase with Dye1600 UnitsContact us
PB80.41-08IsoFast® Hot Start Bst Polymerase with Dye8000 UnitsContact us
PB80.42-01IsoFast® Hot Start Bst Mix100 x 25 μL ReactionsContact us
PB80.42-05IsoFast® Hot Start Bst Mix500 x 25 μL ReactionsContact us

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More Information

IsoFast® Hot Start Bst Polymerase & Mix represents the latest development and improvement in isothermal amplification technology, designed to deliver unmatched specificity and speed in DNA amplification processes and to meet the requirements in molecular diagnostics and research. Whether you are working with Loop-mediated Isothermal Amplification (LAMP) or other isothermal amplification methods, IsoFast® Hot Start Bst Polymerase 2x Mix ensures excellent sensitivity and reliability. Our reagents leverage the power of hot-start Bst DNA polymerase enzyme, ensuring minimal background amplification and maximizing target specificity. With IsoFast® Hot Start, your isothermal amplification experiments will be faster, more efficient, and easier than ever, setting new standards for molecular biology techniques in both diagnostic applications and research efforts.

Hot start is crucial in isothermal amplification techniques because it helps prevent non-specific amplification and false positive results. Isothermal amplification methods, such as 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 important in isothermal amplification?

Preventing non-specific amplification in isothermal Bst polymerase reactions:

In isothermal amplification, it is important to ensure that the amplification process targets only the specific sequence you are interested in. Non-specific amplification can occur when primers bind to unintended regions of the sample or when amplification enzymes initiate too early. Hot start techniques involve inhibiting the amplification reaction until the reaction reaches the desired temperature. This prevents the formation of non-specific products in the early stages of the reaction when the temperature is still below the optimal amplification temperature.

Enhancing specificity of isothermal amplification by Bst polymerase:

By blocking early amplification, hot start methods increase the specificity of the isothermal amplification reaction. This means the reaction is less likely to amplify non-target sequences, reducing the risk of false positive results.

Improving sensitivity in isothermal amplification with Bst polymerase:

Hot start techniques can also improve the sensitivity of isothermal amplification assays. By minimizing non-specific amplification, more reagents are available for amplifying the target sequence, enhancing the efficiency of the reaction and the limit of detection.

Applications

  • Whole Genome Amplification
  • Multiple Displacement Amplification
  • Circular Amplification
  • Isothermal Amplification
  • Loop-mediated Isothermal Amplification (LAMP)
  • Molecular Diagnostics
  • Point-of-care Testing

Specifications

IsoFast® Hot Start Bst Polymerase

Component 1600 Units 8000 Units
IsoFast Hot Start Bst Polymerase 8 U/μL 1 x 200 μL 1 x 1 mL
10x IsoFast Buffer A 1 x 500 μL 2 x 1.25 mL
5x IsoFast Buffer B 1 x 1 mL 3 x 1.7 mL

IsoFast® Hot Start Bst Polymerase with Dye

Component 1600 Units 8000 Units
IsoFast Hot Start Bst Polymerase 8 U/μL 1 x 200 μL 1 x 1 mL
10x IsoFast Buffer A 1 x 500 μL 2 x 1.25 mL
5x IsoFast Buffer B 1 x 1 mL 3 x 1.7 mL
20x Fluorescent Dye 2 x 125 μL 2 x 625 μL

IsoFast® Hot Start Bst Mix

Component 100 Reactions 500 Reactions
2x IsoFast Hot Start Bst Mix 1 x 1.25 mL 5 x 1.25 mL
20x Fluorescent Dye 1 x 125 μL 1 x 625 μL

Reaction Information

Reaction Volume Storage
25 μL Upon arrival, it is recommended to store the product at -30 to -20 °C. If stored properly, the kit will retain activity until the specified expiration date.

Frequently Asked Questions (FAQs)

Certain isothermal amplification strategies can be used for downstream applications, for example, RCA (rolling circle amplification) or WGA (whole genome amplification) for NGS. IsoFast® Hot Start Bst reagents can be used for this type of amplification. However, the most common form of isothermal amplification, LAMP, produces a variety of DNA products, making it unsuitable for cloning and impractical for NGS applications.

The IsoFast® Hot Start Bst Polymerase enzyme, IsoFast® Hot Start Bst Mix, IsoFast® Hot Start Bst Colour Mix, and IsoFast® Hot Start Bst Polymerase Colour are all designed to amplify DNA and cDNA targets. They are not suitable for direct amplification of RNA targets.

For amplifying RNA targets, we recommend using our IsoFast® Bst 1-Step Mix for one-step isothermal amplification. Alternatively, we suggest a two-step format where RNA is first converted to cDNA using one of our UltraScript® reagents, followed by cDNA amplification using the user's preferred IsoFast® Hot Start Bst reagents.
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