SupremeNZYTaqIIDNA_pol

Supreme NZYTaq II DNA polymerase (MB355)

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Description: Supreme NZYTaq II DNA polymerase is an engineered version of NZYTaq II DNA polymerase displaying a hot-start-like PCR capacity. The enzyme activity at room temperature is limited, avoiding extension of non-specifically annealed primers or primer-dimers and thus providing higher specificity, sensitivity and yield during DNA amplification. The functional activity of the enzyme is restored during a short 5-minute incubation step at 95 °C. Once Supreme NZYTaq II DNA polymerase is activated, it displays high performance leading to high DNA yields in shorter extension times (15-30 s/kb) of DNA templates up to 6 kb. Supreme NZYTaq II DNA polymerase requires minimal optimization. In addition, the hot-start-like capacity of Supreme NZYTaq II DNA polymerase not only leads to higher PCR sensitivity but also allows a room-temperature reaction setup. Supreme NZYTaq II DNA polymerase lacks 3´→5´ exonuclease activity. Thus, resulting PCR products have an A-overhang and are suitable for cloning with NZY-A PCR cloning kit (MB053) or NZY-A Speedy PCR cloning kit (MB137).

Features:
– Reproducible and convenient (provides the convenience of room temperature reaction set-up)
– 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
– Amplification of low-copy templates
– High specificity amplifications
– Hot-start PCR
– Genotyping
– Generation of products for TA cloning

Specifications:

Product length:0-6 kb
Fidelity (vs. Taq):
Hot-start-like capacity:yes
Extension time:15-30 sec/kb at 72 °C
3´→5´ activity:no
Product overhang:3’-A
Available as Mixes:Supreme NZYTaq II 2x Green Master Mix (MB360); Supreme NZYTaq II 2x Colourless Master Mix (MB359)
Storage conditions:Store at -20 ºC
Shipping conditions:Shipped at 4 ºC to dry ice

Components:
– Supreme NZYTaq II DNA polymerase (5 U/μL)
– Reaction buffer (10x)
– MgCl2 Solution (50 mM)

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 ° 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) Inadequate PCR Setup. Adjust the primers concentration. Be sure to add the DNA template in last, and that you prepare the PCR mix at room temperature.

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. Adjust concentration of magnesium. Generally, 2-3 mM MgCl2, typically 2.5 mM final concentration, works well for the majority of PCR reactions. Optimal concentration depends on target template, buffer and dNTPs. Optimise magnesium concentration by supplementing MgCl2 in 0.5 increments up to 4 mM.

3. How should I store my Supreme NZYTaq II DNA polymerase?
The product should be stored at -20 ºC, in a constant temperature freezer.

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