NeoTaqII DNA polymerase

Description

The new generation of Taq-derived DNA polymerases NB-60-0005 optimized for standard PCR applications.
Engineered to produce high DNA yields in shorter PCR running times (15-30 s/kb extension).
Lacks 3´→5´ exonuclease activity and supports reliable amplification of a wide range of DNA templates up to 6 kb.
Suitable for PCR under minimal optimization conditions.

Storage Conditions

All components should be stored at -20°C in a freezer without defrost cycles to ensure maximal shelf life.
The enzyme remains stable at 4°C or room temperature for up to 3 days without compromising its stability.
The product will remain stable until the expiry date if stored as specified.

Unit Definition

One unit of the enzyme is defined as the amount required to catalyze the incorporation of 10 nmoles of dNTPs into acid insoluble material in 30 minutes at 72 °C under controlled assay conditions.

Enzyme Concentration

Magnesium Chloride Solution

A provided 50 mM MgCl2 solution allows users to optimize the Mg2+ concentration in PCR setups.
A concentration of 2.5 mM MgCl2 is generally effective with Neo Biotech Taq II DNA polymerase.
Thoroughly vortex the MgCl2 solution after thawing.

Primer Design

PCR primers should range from 15-30 bases and flank the region of interest.
Primers should contain 40-60% GC content and avoid sequences that may produce internal secondary structure.
The 3'-ends of the primers should not be complementary to avoid primer-dimer formation.
Avoid three consecutive G or C nucleotides near the 3'-end to prevent non-specific primer annealing.
Ideally, both primers should have nearly identical melting temperatures (Tm) for efficient annealing.

DNA Template

The recommended amount of genomic DNA template is 10 ng to 500 ng, but as little as 5 pg can be used.
Lower amounts may be sufficient for amplification of less complex DNA (1-20 ng).
When using cDNA synthesis as a template, do not exceed 10% of the final PCR reaction volume.

Quality Control Assays

Taq II DNA polymerase is tested for purity, with >90% purity determined by SDS polyacrylamide gel electrophoresis and Coomassie Blue staining.
Genomic DNA contamination is evaluated through PCR and must not be detectable.

Nuclease Assays

Nuclease assays are performed using pNZY28 plasmid DNA and Neo Biotech Taq II DNA polymerase, followed by visualization on a GreenSafe Premium-stained agarose gel.
No visible nicking or cutting of the nucleic acid should be observed. Similar tests are conducted with the reaction buffer and MgCl2 solution.

Functional Assay

Taq II DNA polymerase is extensively tested for performance in PCR amplification of different-sized DNA fragments from human genomic DNA.
The resulting PCR products should appear as single bands on a stained agarose.