IsoPol® SD+
IsoPol® SD+ is a DNA polymerase with stronger strand displacement and higher salt tolerance, compared to ArcticZymes first IsoPol™.
Figure 1: Temperature optimum of IsoPol® SD+. The enzyme was assayed at the indicated temperatures in 10 mM Tris-HCl pH 8.5, 100 mM NaCl, 5 mM MgCl2, 700 μg/mL denatured herring sperm DNA, 33 μM dNTPs (each), 0.125 μM [3H]-dTTP, 0.8 mM DTT, 0.02% Triton X-100. Samples were incubated for 1 and 5 minutes. Activity was determined measuring radioactive decay [CPM] of incorporated [3H]-dTTPs in acid insoluble material. ΔCPM/min between time point one and two were used as measure of activity. Results are depicted as relative to maximum. The dashed line represents a stipulation of curve progression based on IsoPol® DNA Polymerase
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 37°C. IsoPol® SD+ exhibits a specific activity of ~7 500 U/mg.
Strand-displacement activity: IsoPol® SD+ exhibits stronger strand-displacement activity than other polymerases at 25°C and 37°C.
Features: IsoPol® SD+ exhibits 5’-3’ polymerase activity and lacks 3’-5’ exonuclease activity. The 5’-3’ exonuclease domain is truncated, thus the enzyme also lacks 5’-3’ exonuclease activity.
Inactivation: IsoPol® SD+ can be heat inactivated at temperatures above 50°C.
Optimal reaction conditions: Optimal performance is achieved in a pH 8.5 buffer supplemented with 4-6 mM MgCl2 and 100-350 mM salt (NaCl or KCl), at 37°C.
Storage and Stability
The enzyme is stable at -20°C for 3 years in the supplied storage buffer. The enzyme tolerates at least four freeze-thaw cycles (-80°C) without loss of activity. Storage of the enzyme at 200 mM NaCl is recommended for high concentration samples.
Figure 2: Polymerase activity of IsoPol® DNA Polymerase and IsoPol® II as function of NaCl concentrations in 10 mM Tris-HCl pH 8.5 (@ 25°C), 5 mM MgCl2, 0.72 µg/µl denatured herring-sperm DNA, 33 µM dNTPs (incl. [3H]-dTTPs), 0.4 µg/ml IsoPol or IsoPol II (6.5 nM). Incubation temperature/time: 37°C/10 min. Activity determination by counting decay events of incorporated [3H]-dTTPs in acid insoluble DNA product in counts per minute [CPM]. Data represented relative to highest activity of each polymerase. Error bars represent 1x standard deviation of biological replicates.
Quality Control
ArcticZymes is dedicated to the quality of our products. IsoPol® SD+ is manufactured at our ISO 13485 certified facility in Norway.
dsDNA endonuclease activity
25 U IsoPol® SD+ 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® SD+ 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 data and information relating to IsoPol® SD+. The Product is an Early Access Product, and further product information will be generated continuously. Available data includes; buffer and storage conditions, optimal conditions, activity processivity, and strand displacement data.
We appreciate suggestions for product modifications.
Disclaimer:
These products are intended for further manufacturing use or research use only. Certain applications of ArcticZymes AS products may require licenses from others. It is the expressed duty of any receiver of ArcticZymes AS products to acquire such licenses, if necessary. To the extent allowed by law, ArcticZymes AS will not be liable for damages, whether direct, indirect, incidental, or consequential in connection with or arising from this document, including the use of it. ArcticZymes AS products may be covered by pending or issued patents, designs or design applications and/or trademarks or trademark applications or any other registered or unregistered Intellectual Property Right.