Speedy NZYProof DNA polymerase (MB404)

62,00226,00

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Description: Speedy NZYProof DNA polymerase belongs to a new generation of DNA polymerases displaying a fast polymerization rate. Only 5 seconds are required for the successful synthesis of a 1 kb size DNA fragment. Faster PCR can be further achieved by increasing the primers melting temperature, which increases PCR annealing temperature, thus allowing combining the annealing and extension PCR steps (see below). Speedy NZYProof DNA polymerase is supplied with an optimized 5× Reaction Buffer and a 50 mM MgCl2 solution. The enzyme supports robust and reliable reactions while tolerating a wide range of templates. It is useful for fast and routine DNA amplifications thar require fidelity, as well as for genotyping. PCR products generated by Speedy NZYProof DNA polymerase have blunt-ends and are suitable for cloning with NZYTech´s NZY-blunt PCR cloning kit (NZYTech, Cat. No. MB121).

Features:
– Fast amplification (5 sec/kb)
– Proofreading activity

Applications
– High-fidelity PCR
– Fast PCR
– Generation of products for blunt-end cloning

Successful amplification with limited extension time and proofreading activity:
Three different DNA fragments of various sizes and from different origins were amplified using a limited extension period of 5 sec/kb and 2.5 U of Speedy NZYProof DNA polymerase.

speedyproof Amplification of nucleic acids from different DNA templates using Speedy NZYProof DNA polymerase. Lane 1: 350 bp PCR product (sheep genomic DNA, 120 ng). Lane 2: 1 kb PCR product (E. coli genomic DNA, 100 ng). Lane 3: 2 kb PCR product (plasmid DNA, 10 ng). Lane M: NZYDNA Ladder III.

Specifications:

Product length: 0-3 kb
Hot-start like capacity: no
Speed: 5 sec/kb
Product overhang: blunt
Storage conditions: Store at -20 ºC
Shipping conditions: Shipped at 4 ºC to dry ice

Components:
– Speedy NZYProof DNA polymerase (2.5 U/μL)
– Reaction buffer (10x)
– MgCl2 (50 mM)

Product Brochure EN
Safety Information EN
Informacao de Seguranca PT
Certificate of Analysis ZN011
Certificate of Analysis YL121
Certificate of Analysis NM101

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.

2. How should I store my Speedy NZYProof DNA polymerase?
The product should be stored at -20 ºC, in a constant temperature freezer.

3. I’d like to clone a fragment amplified with NZYSpeedy Proof DNA polymerase. How should I proceed?
Speedy NZYProof DNA polymerase has 3´→ 5´ exonuclease activity and will generate blunt-ended products. So, blunt-end cloning is recommended, by using NZY-blunt PCR cloning kit (MB121) or other similar cloning kit. Nevertheless, if TA cloning is required, the generated blunt-ended PCR fragments can be modified by adding 3´A-overhangs with a different polymerase, such as Taq DNA polymerase (see the A-tailing protocol in the product brochure of NZY-A PCR cloning kit – MB053). Due to the strong proofreading activity of Speedy NZYProof DNA polymerase, we recommend immediately proceed with cloning after this procedure, in order to avoid lost of the 3´A-overhangs during storage. Alternatively, the PCR products can be cloned using the innovative NZYEasy Cloning & Expression Systems in a rapid, precisely and ligase-independent manner. For optimal cloning efficiencies, silica-column purification of the PCR product using NZYGelpure kit (MB011) or other similar kit is highly recommended. This procedure will remove dNTPs, unused primers and other impurities. Spin-column purification is sufficient as long as the product is >90% pure. Gel-extraction should be performed in case non-specific amplifications and/or primer-dimers are formed.

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