ArcticZymes (Stripe Checkout Test)

OverView

AZtaq™ DNA Polymerase is a high-quality DNA polymerase, originating form Thermus aquaticus. Being highly thermostable, AZtaq is ideal for use in polymerase chain reaction (PCR) applications.

The enzyme catalyses the synthesis of a complementary DNA strand using a primed DNA or cDNA strand as template. It possesses 5’-3’ exonuclease activity while lacking 3’-5’ proofreading activity.

AZtaq is compatible with the use of dUTP, enabling highly efficient removal of carry-over contamination with Cod UNG.

Key Features

Excellent qPCR Performance
Compatible with dUTP
Thermostable

Applications

PCR/qPCR

Figures

Figure 1. AZtaq shows excellent performance and sensitivity in qPCR

Figure 1.2. AZtaq shows excellent performance and sensitivity in qPCR

A serial dilution of M13KO7 DNA was used as template for qPCR amplification using specific primers together with a fluorescently labelled hydrolysis probe detecting the M13v1.1 target. Data showing amplification plots (A) and standard curve (B) for the qPCR reactions containing from 1 000 000 down to 1 copy of M13v1.1 target DNA, in triplicate measurements.

Properties

Source

Recombinantly produced in E. coli

Size

95.1 kDa

Unit Definition

One unit (U) is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 min at 72°C. The enzyme is assayed in 20 mM Tris-HCl pH 8.8, 55 mM KCl, 2 mM MgCl2, 0.02 mg/ml BSA, 15 nM primed M13mp18 ssDNA, 0.8 mM dNTPs (0.2 mM each), 1.888 µM 3H-dTTP.

Inactivation

N/A

Storage and Stability

AZtaq is stable at -20°C for up to 1 year in the supplied storage buffer. Additional data on stability is available on request.

Quality Control

ArcticZymes is dedicated to the quality of our products. AZtaq is manufactured at our ISO 13485 certified facility in Norway.

dsDNA endonuclease activity

20 U AZtaq 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

20 U AZtaq 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.

dsDNA exonuclease activity

20 U AZtaq was incubated with 3H-dATP labelled dsDNA (0.5 µg, 500 bp) for 4 hrs at 37°C. Acid soluble radioactivity from labelled DNA was not significantly over blank test for either substrate.

RNase activity

20 U AZtaq was incubated with a 2 kb RNA transcript (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of RNA degradation.

E. coli gDNA

5 U AZtaq was analysed in a probe-based qPCR assay detecting the 23S ribosomal RNA gene in E. coli. Less than 3 copies of E. coli gDNA were detected.

Functional test

5 Units AZtaq was used in a qPCR assay amplifying a serial dilution of down to < 3 copies of E. coli gDNA using specific primers detecting the 23S ribosomal RNA gene. Successful amplification was observed for all dilutions. The resulting standard curve showed at least 90-110 % efficiency, indicating efficient doubling of specific template product per cycle.