Ultra-High Fidelity Including dNTP's RadiantTM HiFi Ultra DNA Polymerase is a high-performance, high-fidelity DNA Polymerase optimized for the sensitive DNA amplification of a wide range of DNA templates including complex mammalian genomic DNA. RadiantTM HiFi Ultra Polymerase possesses 5´-3´ DNA polymerase and 3´-5´ proofreading exonuclease activities with an error-rate of 4.55 x 10-7 . The polymerase is ideal for cloning applications, library construction, screening and genotyping of problematic GC-rich and AT-rich sequences. RadiantTM HiFi Ultra Polymerase is engineered for robust, superior high-fidelity PCR in comparison with standard proof-reading enzymes such as pfu DNA Polymerase. Several point-mutations combined with a new-generation buffer formulation provide for significantly improved processivity for faster cycling, greater sensitivity and less inhibition with crude DNA samples. The supplied 5X HiFi Ultra Reaction Buffer contains an ideal concentration of MgCl2 and highly pure dNTPS for convenience and consistency in liquid handling. Storage RadiantTM HiFi Ultra DNA Polymerase is shipped on blue or dry ice and should be stored at –20°C upon receipt. Excessive freeze/thawing should be avoided. When stored as specified, RadiantTM HiFi Ultra DNA Polymerase is stable for 12 months from date of receipt. The Kit may also be stored at 4°C for 1 month. Important Considerations RadiantTM 5x HiFi Ultra Reaction Buffer: The 5x reaction buffer contains 15mM MgCl2, 5mM dNTPs, and PCR enhancers. The buffer composition has been optimized for maximum efficiency, sensitivity and success with difficult amplicons. We do not suggest the use of additional MgCl2 or enhancers. Template: For complex genomic DNA, we suggest the use of 5ng - 500ng per reaction; For cDNA or plasmid DNA, please use < 100ng per reaction. Primers: Primers should have a predicted melting temperature of around 60°C, using default Primer 3 settings (http://frodo.wi.mit.edu/primer3/). The final primer concentration in the reaction should be between 0.2μM and 0.6μM. Annealing: We recommend performing a temperature gradient to determine the optimal annealing temperature. Alternatively, we suggest a 57°C annealing temperature and then an increase in 2°C increments if non-specific products are present. Extension: Optimal extension is achieved at 72°C. The optimal extension time is dependent on amplicon length and complexity. 30 seconds per kilobase(Kb) is recommended for amplification from eukaryotic genomic DNA or cDNA. Quality Control RadiantTM HiFi Ultra DNA Polymerase is tested extensively for robust activity, processivity, efficiency, heat activation, sensitivity, absence of nuclease contamination and absence of nucleic acid contamination. RadiantTM HiFi Ultra DNA Polymerase is manufactured under a comprehensive quality management system, following ISO 9001:2008 standards. Limitations of Use This product is intended for research purposes only and is not intended for any animal or human therapeutic use. Technical Support For Trouble-shooting and Technical Guidance, please contact us at tech@alkalisci.com and provide PCR reaction conditions, cycling parameters, amplicon size, and screen grabs (gel images) if possible. Alkali Scientific guarantees the performance of all products in the manner described in our product literature. The purchaser must determine the suitability of the product for its particular use. Alkali Scientific shall not be liable for any direct, indirect, consequential or incidental damages arising out of the use, the result of use or the liability to use this product.