NZYLong 2x Green Master Mix (MB139)


Description: NZYLong 2× Green Master Mix is a premixed ready-to-use solution containing NZYLong DNA polymerase (MB003), an engineered DNA polymerase designed to amplify long target DNA sequences, generally of 20 kb and beyond. A wide range of long PCR products can be generated using lambda DNA or human genomic DNA as starting template. The master mix contains dNTPs, reaction buffer and additives at optimal concentrations for the efficient amplification of an extensive 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 NZYLong 2× Green Master Mix may be directly loaded onto agarose gels. There are two dyes (blue and yellow) in the mix that allow monitoring the progress of electrophoresis.

NZYLong DNA polymerase generates a mixture of A-overhang-ended (predominantly) and blunt-ended PCR products, being suitable for cloning with NZYTech´s TA PCR cloning kits (MB053 or MB137).

– Convenient ready-to-use PCR master mix
– Direct Gel Loading
– Amplifies long DNA fragments (up to 20 kDa)
– Higher fidelity than NZYTaq DNA polymerase

– Long-Range PCR
– Colony PCR
– Generation of products for TA cloning

Convenience of direct gel loading of different-sized PCR products:
The convenience of NZYLong 2x Green Master Mix can be demonstrated through an high-throughput PCR experiment with 96 genes of different sizes (from around 0.5 to around 5 kb).



Product length: 0-20 kb
Hot-start-like capacity: no
Extension time: 60 sec/kb at 68 °C
3´→5´ activity: no
Product overhang: mixed
Storage conditions: Store at -20 ºC
Shipping conditions: Shipped at room temperature to dry ice

NZYLong 2× Green Master Mix (0.2 U/μL)

Product Brochure EN
Safety Information EN
Informacao de Seguranca PT
Certificate of Analysis ZN071

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) Template DNA damaged or degraded. An intact, high-quality template is essential to achieve a reliable amplification of large DNA fragments. Extreme care must be taken in the preparation and handling of DNA. Always use purified high-quality DNA as template.
d) 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. Check the quality and concentration of primer solutions. We recommend to prepare small-volume working aliquots from the stock solution.  Avoid using primers subjected to multiple freezing-thawing cycles.

3. How should I store my NZYLong 2× Green Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.

Next-generation sequencing and comparative analysis of Pyrrhocorax pyrrhocorax and Pyrrhocorax graculus (Passeriformes: Corvidae) mitochondrial genomes
Morinha F, Clemente C, Cabral JA, Lewicka MM, Travassos P, Carvalho D, Dávila JA, Santos M, Blanco G, Bastos E.
Mitochondrial DNA A DNA Mapp Seq Anal, 2016



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