IsoPol™ BST+

Overview

IsoPol™ BST+ is a heat-tolerant Bst polymerase (large fragment) with enhanced strand-displacement activity. IsoPol™ BST+ is active from 25 to 65°C. It is lacking 5’-3’- and 3’-5’-exonuclease activity.

Properties

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

Strand-displacement activity: IsoPol™ BST+ exhibits enhanced strand-displacement activity compared to Bst polymerase.

Strand displacement activity with increasing concentrations of KCI

Enhanced strand displacement activity compared to Bst polymerase
Figure 1: IsoPol™ BST+ shows enhanced strand displacement activity compared to Bst polymerase. Activities at different concentrations of KCl at 37°C relative to activity at 50 mM KCl are shown for each polymerase.
Tolerates higher concentrations of salt compared to Bst polymerase
Figure 2: IsoPol™ BST+ tolerates higher concentrations of salt compared to Bst polymerase. Polymerase activity of Bst and IsoPol™ BST+ as function of NaCl data represented relative to highest activity of each polymerase.

Features: IsoPol™ BST+ lacks 5’-3’- and 3’-5’-exonuclease activity.

Inactivation: IsoPol™ BST+ is completely inactivated after one minute incubation at 95°C

Suggested reaction conditions: IsoPol™ BST+ is active at pH 8.5 in buffer supplemented with 5-20 mM MgCl2 and 50-350 mM salt (NaCl or KCl), at 65°C.

Storage and Stability

The enzyme is stable at -20°C for > 1 year in the supplied storage buffer. The enzyme tolerates at least four freeze-thaw cycles (-80°C) without loss of activity.

Quality Control

ArcticZymes is dedicated to the quality of our products. IsoPol™ BST+ is manufactured at our ISO 13485 certified facility in Norway.

dsDNA endonuclease activity

25 U IsoPol™ BST+ was incubated with a supercoiled plasmid (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any transformation of closed circular DNA to nicked DNA.

ssDNA endonuclease activity

25 U IsoPol™ BST+ was incubated with M13 ssDNA (0.5 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of ssDNA degradation.

Additional Data and Information

We are pleased to provide additional data relating to IsoPol™ BST+ upon request.

We appreciate suggestions for product modifications.

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