Cat. No.: BSP-8k (for 8KU)

Cat. No.: BSP-40k (for 40KU)

For better isothermal amplification of RNA, please see Bst DNA/RNA Polymerase.

For custom-ready mastermix (including visualization dye, probes, primers, etc), please email your requirements to tech@sbsbio.com.

The glycerol-free version is also available for this product, which can be used to establish a freeze-drying system.

Description

Bst Polymerase is a homologous protein of Bst DNA Polymerase, large fragment, derived from the thermophilic bacterium Bacillus stearothermophilus (Bst). When compared to the large fragment of Bst DNA Polymerase, it exhibits stronger 5'→3' DNA polymerase activity, enhanced strand displacement capability, tolerance to dUTP, salt resistance, and resistance to non-ionic detergents. Unlike Bst DNA Polymerase large fragment, Bst Polymerase lacks 5'→3' and 3'→5' exonuclease activity. It can be used in various applications, including loop-mediated isothermal amplification (LAMP), crossing priming amplification (CPA), rolling-circle amplification (RCA), and isothermal amplification reactions based on rolling-circle amplification.

The isothermal amplification temperature mediated by Bst Polymerase generally falls between 50-72°C, typically at 65°C. The optimal temperature depends on the primers and the products being amplified and may require experimental optimization.

Compared to the Bst 3.0 DNA Polymerase from similar companies, this product not only exhibits comparable DNA polymerase activity, strand displacement capability, dUTP tolerance, and high-temperature resistance (tolerance up to 72°C), but it also demonstrates superior reverse transcriptase activity and stronger dUTP tolerance. This makes it suitable for reverse transcription loop-mediated isothermal amplification (RT-LAMP).

This product possesses all the excellent features of Bst DNA Polymerase. Compared to Bst DNA Polymerase, it not only exhibits comparable DNA polymerase activity, strand displacement capability, salt resistance, and non-ionic detergent tolerance but also has superior high-temperature tolerance (up to 72°C), better reverse transcriptase activity, and stronger dUTP tolerance.

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10nmol of dNTP into acid insoluble material in 30 minutes at 65ºC.

Source

Bst Polymerase is obtained through recombinant expression and purification in Escherichia coli.

Purity

Free from DNA endonuclease and exonuclease activities.

Applications

Suitable for various DNA isothermal amplification techniques, including loop-mediated isothermal amplification (LAMP), reverse transcription loop-mediated isothermal amplification (RT-LAMP) (reaction temperature can reach 72°C), crossing priming amplification (CPA), rolling-circle amplification (RCA), helicase-dependent amplification (HDA), multiple displacement amplification (MDA), strand displacement DNA synthesis, whole-genome amplification (WGA), high GC-content DNA sequencing, rapid sequencing of nanogram-level DNA templates, library sequencing, and more.

Enzyme Storage Buffer

10mM Tris-HCl, 50mM KCl, 0.1mM EDTA, 1mM DTT, 0.1% Triton X-100, 50% Glycerol (pH 7.5 @ 25°C).

10X Reaction Buffer

200mM Tris-HCl, 500mM KCl, 100mM (NH4)2S04, 20mM MgS04, 1% Tween-20 (pH 8.8 @ 25°C).

Inactivation or Inhibition

Bst Polymerase can be inactivated by heating at 80°C for 5 minutes.

Storage Conditions

Store the components at -20°C. Avoid multiple freeze-thaw cycles.

Instruction: Protocol

Related: 

• Bst DNA Polymerase Large Fragment
• Bst DNA Polymerase (comparable to 2.0)
• Bst Polymerase (comparable to 3.0)
• Bst Polymerase (glycerol-free)
• Bst DNA/RNA Polymerase (with superior reverse transcription activity) 
• Bst DNA/RNA Polymerase (glycerol-free)
• Bst P DNA/RNA Polymerase (glycerol-free) (with superior reverse transcription activity & hot-start property) 

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Only for research and not intended for treatment of humans or animals