Description: Supreme NZYProof DNA polymerase is an engineered highly accurate, fast and sensitive variant of NZYProof DNA polymerase displaying a hot-start like PCR capacity. This feature is achieved by a novel hot-start technology, which inhibits both polymerase and 3’→5’ exonuclease activities and thus avoids extension of non-specifically annealed primers or primer-dimers, as well as the degradation of primers and template DNA during PCR reaction setup. Thus, Supreme NZYProof DNA polymerase generates higher specificity, sensitivity and yield during the accurate amplification of DNA. Supreme NZYProof DNA polymerase was also engineered for higher processivity, thus allowing fast PCR reactions of longer PCR products.
This highly robust version of NZYProof DNA polymerase is a broad range enzyme suitable for a variety of applications, including amplification of longer (≤10 kb) and difficult PCR products, as well as site-directed mutagenesis. Supreme NZYProof DNA polymerase possesses 3´→5´ exonuclease proofreading activity that enables the polymerase to amplify DNA with increased accuracy. The error rate of Supreme NZYProof DNA polymerase is similar to that of Pfu and Kod DNA polymerases and significantly lower than the error rate of Taq DNA polymerases. Supreme NZYProof is 50x more reliable than NZYTaq DNA polymerase (standard polymerase). It generates blunt-ended PCR products that are suitable for cloning with NZYTech´s NZY-blunt PCR cloning kit (MB121).
Features:
– High sensitivity
– Eliminates nonspecific amplifications and minimizes primer-dimers
– High processivity/ Fast amplification (15-30 sec/kb)
– Proofreading activity
Applications:
– High-fidelity PCR
– Hot-start PCR
– Amplification of difficult-to-amplify targets (e.g. high GC-contents)
– Site-directed mutagenesis
– Generation of products for blunt-end cloning
High specificity hot-start PCR
Supreme NZYProof DNA polymerase was engineered to exhibit extreme specificity, enabling accurate amplification of a broad range of DNA templates with different sizes and concentrations. During reaction setup, Supreme NZYProof DNA polymerase is inactivated through a novel hot-start technology, which inhibits both polymerase and 3’→5’ exonuclease activities of the enzyme at room temperature, as well as on ice. Supreme NZYProof DNA polymerase does not require a separate activation step during the PCR cycling, since its activity is immediately restored at high temperatures.
Three E. coli genomic DNA fragments of different lengths (1, 2.5 and 5 kb) were amplified with Supreme NZYProof DNA polymerase (2.5 U). Three 4-fold gDNA dilutions ranging from 40 to 2.5 ng were used as starting template to amplify each DNA fragment. NC: No template control. Lane M: NZYDNA Ladder III (MB044). Results demonstrate amplification in all the conditions tested with high yields and specificity.
Specifications:
| Product length: | 0-10 kb |
| Hot-start like capacity: | yes |
| Extension time: | 15-30 sec/kb at 72 ºC |
| 3´→5´ activity: | no |
| Product overhang: | blunt |
| Available as Mixes: | Supreme NZYProof 2x Green Master Mix (MB285); Supreme NZYProof 2x Colourless Master Mix (MB286) |
| Storage conditions: | Store at -20 ºC |
| Shipping conditions: | Shipped at 4 ºC to dry ice |
Components:
– Supreme NZYProof DNA polymerase (2.5 U/μL)
– Reaction buffer (5x)
