Speedy Supreme NZYTaq 2x Colourless Master Mix (MB392)

99,00842,00

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Description: Speedy Supreme NZYTaq 2× Colourless Master Mix is a premixed ready-to-use solution containing Speedy Supreme NZYTaq DNA polymerase (MB390), 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. The master mix contains dNTPs, reaction buffer and additives at optimal concentrations for the efficient amplification of a wide range of DNA templates. MgCl2 final concentration is 2.5 mM, allowing the implementation of a variety of PCR protocols.

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:
– Convenient ready-to-use PCR master mix
– 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
– Colony PCR
– Generation of products for TA cloning

Excellent performance of Speedy Supreme NZYTaq 2x Colourless Master Mix in fast amplification of diferent-sized DNA fragments
The performance of Speedy Supreme NZYTaq 2x Colourless Master Mix was tested in a fast PCR assay to amplify fragments of 1 kb, 2.5 kb and 5kb from diferent dilutions of human genomic DNA.SpeedySupremeNZYTaqMix

The protocol provided with the NZYTech’s Master Mix 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. A 5-fold human genomic DNA dilution, ranging from 20 to 0.032 ng, was used for the amplification of  diferent-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
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
Storage conditions: Store at -20 ºC
Shipping conditions: Shipped at 4 ºC to dry ice

Components:
Speedy Supreme NZYTaq 2× Colourless Master Mix (0.2 U/μL)

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 °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) 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.

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

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