Speedy Supreme NZYTaq 2x Green Master Mix (MB391)

99,00842,00

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Description: Speedy Supreme NZYTaq 2× Green Master Mix is a premixed ready-to-use solution containing Speedy Supreme NZYTaq DNA polymerase (MB390), a recombinant modified form of Taq DNA polymerase that combines hot-start like PCR capacity and a fast polymerization reaction. The enzyme is inactive at room temperature, avoiding extension of non-specifically annealed primers or primer-dimers and thus providing higher specificity and sensitivity of DNA amplification. The functional activity of the enzyme is restored during a short 5- minute incubation step at 95 °C. The master mix contains dNTPs, reaction buffer and additives at optimal concentrations for the efficient amplification of a wide range of DNA templates. MgCl2 final concentration is 2.5 mM, allowing the implementation of a variety of PCR protocols. In addition, reactions assembled with Speedy Supreme NZYTaq 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
– Fast routine amplification (5 sec/kb for DNA fragments up to 2-3 kb)
– Eliminates nonspecific amplifications and minimizes primer-dimers
– Robust amplification using a wide range of DNA templates
– Leaves an A-overhang

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

Specifications:

Product length: 0-6 kb
Hot-start-like capacity: yes
Extension time: 5 sec/kb (or 10 sec/kb for targets higher than 2-3 kb in size) 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:
Speedy Supreme NZYTaq 2× Green Master Mix (0.2 U/μL)

MSDS EN
MSDS PT
Product Brochure EN

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 Speedy Supreme NZYTaq 2× Green Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.

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