Bst DNA polymerase, Large Fragment 8,000 units/ml

Description

Bst DNA Polymerase, Large Fragment is a highly thermostable enzyme derived from Bacillus stearothermophilus DNA Polymerase. This recombinant enzyme is expressed in E. coli and engineered to retain the essential 5´→ 3´ polymerase activity while lacking 5´→ 3´ exonuclease activity. The enzyme exhibits exceptional strand displacement capabilities, making it the gold standard for isothermal amplification techniques. 

Features

• Superior Strand Displacement Activity – ideal for isothermal amplification (e.g. LAMP). 
• Optimal Thermostability – works well at 60–65°C.
• Exonuclease Deficient – Lacks both 5´ → 3´ and 3´ → 5´ exonuclease activities, preserving DNA template integrity and preventing degradation of amplification products. 
• High processivity – synthesizes long DNA efficiently.
• Versatile use – suitable for LAMP, MDA, and more.

Applications

• Loop-mediated isothermal amplification (LAMP) 
• Multiple Displacement Amplification (MDA) 
• Rolling Circle Amplification (RCA)
• Whole Genome Amplification (WGA)

Unit Definition

One unit of enzyme activity is defined as the amount of enzyme that incorporates 10 nmol of dNTP into acid-insoluble material within 30 minutes at 65°C.

Storage Temperature: -20°C for optimal long-term stability

Storage Buffer:

25 mM Tris-HCl (pH 8.0), 150 mM NaCl, 0.5 mM DTT, 0.5 mM EDTA, 50% Glycerol

Important Handling Notes:

• Avoid repeated freeze-thaw cycles to maintain enzyme activity
• Store at -20°C for maximum stability
• Thaw on ice before use

Specifications

• Expression System: Escherichia coli
• Theoretical Molecular Weight: 67,000 Daltons
• Concentration: 8,000 units/ml
• Heat Inactivation: 80°C for 20 minutes
• 5′ → 3′ Exonuclease: No
• 3′ → 5′ Exonuclease: No
• Strand Displacement: Yes

Quality Control

Each lot undergoes comprehensive testing including:

• Functional activity assays at 65°C 
• Purity analysis by SDS-PAGE (>98%) 
• Endonuclease and RNase contamination testing
• Sterility verification