IsoFast™ Bst Polymerase with Dye – Witec AG

The enzyme offers fast amplification and strong strand displacement capabilities, making it ideal for nucleic acid amplification methods such as whole genome amplification, multiple displacement amplification and isothermal amplification.

IsoFast™ Bst Polymerase is a recombinant form of the large fragment of Bst DNA polymerase, containing 5′-3′ polymerase activity and strand displacement activity.

• Has strand-displacing 5′-3′ polymerase activity
• Lacks 5′-3′ exonuclease activity
• DNA synthesis is performed at a constant temperature
• Operates over a broad temperature range, with an optimum of 65ºC
• Gives rapid and consistent amplification across a wide range of templates
• Supplied with an advanced 2-part buffer system for higher yields under difficult conditions
• 30 minute protocol
• Flexible formats with and without fluorescent dye
• Also available as a 2x mix
• Glycerol-free enzyme

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 65ºC. However IsoFast™ Bst Polymerase works well over a broad temperature range (55ºC to 70ºC, 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 80ºC.

Rapid and consistent results

Designed for fast amplification speed, IsoFast™ Bst Polymerase gives rapid and consistent results across different target sequences (see Figure 1, 2 & 3). The enzyme is provided with an advanced 2-part buffer system to ensure higher yield and performance even under difficult conditions. Amplification can be detected in a number of ways including real-time fluorescence detection and end-point visualisation.

Format flexibility

For added convenience the enzyme is available with separate fluorescent dye (enabling real-time detection with any qPCR thermocycler), and in a 2x mix format for those looking for reduced setup times. The enzyme gives consistent results regardless of the format chosen.

Applications

• Whole genome amplification
• Multiple displacement amplification
• Isothermal amplification
• Loop mediated isothermal amplification (LAMP)
• Molecular diagnostics
• Field diagnostics

This product is not suitable for PCR.

¹ Mead DA, McClary JA, Luckey JA, Kostichka AJ, Witney FR, Smith LM. Bst DNA polymerase permits rapid sequence analysis from nanogram amounts of template. Biotechniques. 1991 Jul;11(1):76-8, 80, 82-87.