Supreme NZYTaq II 2x Green Master Mix (MB360)

84,00688,00

Description: Supreme NZYTaq II 2× Green Master Mix is a premixed ready-to-use solution containing Supreme NZYTaq II DNA polymerase (MB355), a robust DNA polymerase derived from Taq DNA polymerase that was engineered to exhibit high processivity and to provide high PCR sensitivity. The enzyme, displaying a hot-start-like PCR capacity, is inactive at room temperature, avoiding extension of non-specifically annealed primers or primer-dimers and providing a highly specific DNA amplification. The functional activity of the enzyme is restored during a short 5-minute incubation step at 95 °C. In addition, the master mix contains dNTPs, reaction buffer and additives at optimal concentrations for the efficient amplification of a wide range of DNA templates of up to 6 kb. MgCl2 final concentration is 2.5 mM, allowing the implementation of a variety of PCR protocols. In addition, reactions assembled with Supreme NZYTaq II 2× Green Master Mix may be directly loaded onto agarose gels. The mix contains two dyes (blue and yellow) that allow monitoring the progress of the electrophoresis.

Resulting PCR products have an A-overhang and are suitable for cloning with NZYTech´s NZY-A PCR cloning kit (MB053) or NZY-A Speedy PCR cloning kit (MB137).

Features:
– Convenient ready-to-use PCR master mix
– Direct Gel Loading
– High sensitivity
– Eliminates nonspecific amplifications and minimizes primer-dimers
– Robust amplification using a wide range of DNA templates
– Leaves an A-overhang

Applications:
– Powerful PCR
– Colony PCR
– Generation of products for TA cloning

Specifications:

Product length: 0-6 kb
Hot-start-like capacity: yes
Extension time: 15-30 sec/kb at 72 °C
3´→5´ activity: no
Product overhang: 3’-A
Storage conditions: Store at -20 ºC
Shipping conditions: Shipped at 4 ºC to dry ice

Components:
Supreme NZYTaq II 2× Green Master Mix (0.2 U/μL)

Product Brochure EN
Safety Information EN
Informacao de Seguranca PT
Certificate of Analysis ZN051
Certificate of Analysis ZN031
Certificate of Analysis YO101
Certificate of Analysis YO091
Certificate of Analysis YO031
Certificate of Analysis YO021
Certificate of Analysis NM071
Certificate of Analysis NM061
Certificate of Analysis NM031
Certificate of Analysis NM011

1. What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 °C to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.5 mM increments. (Note: MgCl2 is not provided in separate tubes).

2. How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences.

3. How should I store my Supreme NZYTaq II 2× Green Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.

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