Speedy Supreme NZYTaq DNA polymerase (MB390)

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Description: Speedy Supreme NZYTaq DNA polymerase is 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. Speedy Supreme NZYTaq DNA polymerase is supplied with an optimized 10× Reaction Buffer and a 50 mM MgCl2 solution. Minimal optimizations are required for successful DNA amplification. In addition, the hot-start-like capacity of Speedy Supreme NZYTaq DNA polymerase not only leads to higher PCR sensitivity but also allows a room-temperature reaction setup. The enzyme 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)
– 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
– Amplification of low-copy templates
– High specificity amplifications
– Hot-start PCR
– Genotyping
– Generation of products for TA cloning

High sensitive and specific amplification of diferent-sized DNA fragments using a fast PCR protocol
The elongation rate, as well as the specificity, of Speedy Supreme NZYTaq DNA polymerase was tested in a fast PCR assay to amplify fragments of 1 kb, 2.5 kb and 5kb from different dilutions of human genomic DNA using two amounts of enzyme (1 and 0.5 µL). The protocol provided with the Speedy Supreme NZYTaq DNA polymerase was followed using a limited extension period of 5 sec/kb to amplify the 1 kb fragment and 10 sec/kb to amplify both 2.5 kb and 5 kb.

SpeedySupremeNZYTaqA 5-fold human genomic DNA dilution, ranging from 20 to 0.032 ng, was used for the amplification of  different-sized fragments. Lane M: NZYDNA Ladder III. PCRs were performed in 50 µL reaction mixtures with elongation times of 5 sec, 25 sec, and 50 sec for 1kb, 2.5 kb and 5 kb fragments, respectively. Results show successful amplification in conditions under test.

Specifications:

Product length: 0-6 kb
Fidelity (vs. Taq):
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
Available as Mixes: Speedy Supreme NZYTaq 2x Green Master Mix (MB91); Speedy Supreme NZYTaq 2x Colourless Master Mix (MB392)
Storage conditions: Store at -20 ºC
Shipping conditions: Shipped at 4 ºC to dry ice

Components:
– Speedy Supreme NZYTaq 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 Speedy Supreme NZYTaq DNA polymerase?
The product should be stored at -20 ºC, in a constant temperature freezer.

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