NZYSpeedy Proof DNA polymerase (MB109)

59,00219,00

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Description: NZYSpeedy Proof DNA polymerase is a recombinant thermostable DNA polymerase purified from Escherichia coli that combines high fidelity and fast polymerization capacities. NZYSpeedy Proof DNA polymerase possesses 3´→5´ exonuclease proofreading activity that enables the polymerase amplifying DNA with increased accuracy and yield. The error rate of the enzyme is similar to that of Pfu and Kod DNA polymerases (the order of magnitude is between 10-5 and 10-6), which is significantly lower than the error rate of Taq DNA polymerases. NZYSpeedy Proof DNA polymerase is highly efficient in the amplification of DNA fragments up to 3 kb in a short reaction period. Only 15 seconds are required for the successful synthesis of 1 kb size DNA. NZYSpeedy Proof DNA polymerase generates blunt-ended polymerase chain reaction (PCR) products that are suitable for cloning with NZYTech´s NZY-blunt PCR cloning kit (MB121).

Features:
– Fast amplification (15 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 15 sec/kb and 2.5 U of NZYSpeedy Proof DNA polymerase.

speedyproof Amplification of nucleic acids from different DNA templates using NZYSpeedy Proof 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: 15 sec/kb
Product overhang: blunt
Storage conditions: Store at -20 ºC
Shipping conditions: Shipped at 4 ºC to dry ice

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

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 ° 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 NZYSpeedy Proof 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?
NZYSpeedy Proof 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 NZYSpeedy Proof 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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