NZYTaq II with 5x Gel Load Reaction Buffer (MB364)

39,00176,00

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Description: NZYTaq II DNA polymerase belongs to a new generation of Taq-derived DNA polymerases that was engineered to produce high DNA yields in shorter PCR running times (15-30 s/kb extension) under minimal optimization conditions. NZYTaq II DNA polymerase supports the robust and reliable amplification of a wide range of DNA templates up to 6 kb. 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. 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
– Robust amplification using a wide range of DNA templates
– Leaves an A-overhang

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

Specifications:

Product length:0-6 kb
Hot-start-like capacity:no
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 room temperature to dry ice

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

Product Brochure EN
MSDS EN
MSDS PT
Certificate of Analysis NM031

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 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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