Supreme NZYTaq II with 5x Gel Load Reaction Buffer (MB365)

79,00359,00

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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. Supreme NZYTaq II is provided with 5× Gel Load Reaction Buffer allowing reactions to be loaded directly into gels without the extra adding of loading dye. This Gel Load Reaction Buffer is composed by a blue and yellow dye. The blue dye migrates at the same rate as a 3-5 kb DNA fragment in a 1% (w/v) agarose gel. The yellow dye migrates at a rate faster than primers (<50 bp) in a 1% (v/v) agarose gel. The 5× Gel Load Reaction Buffer is not suitable when direct fluorescent or absorbance readings are required without prior purification of the amplified DNA from PCR. Supreme NZYTaq II DNA polymerase lacks 3´→5´ exonuclease activity. 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:
– Suitable for immediate loading onto agarose gels
– High sensitivity and specificity
– Reproducible and convenient (provides the convenience of room temperature reaction set-up)
– Leaves an A-overhang

Applications:
– Powerful PCR
– Amplification of low-copy templates
– High specificity amplifications
– Hot-start 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 ºC
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 DNA polymerase (5 U/μL)
– Gel Load Reaction buffer (5x)
– MgCl2 Solution (50 mM)

MSDS EN
MSDS PT
Product Brochure EN
Certificate of Analysis NM071
Certificate of Analysis NM072

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) Additives required. Adding PCR-enhancing agents, like NZYTaq 5x Optimizer Solution (MB060) or NZYTaq 2x GC-Enhancer solution (MB143) may improve yield while amplifying difficult templates.

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 with 5x Gel Load Reaction Buffer?
The product should be stored at -20 ºC, in a constant temperature freezer.

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