Radiant™ GC-Long DNA Polymerase RadiantTM GC-Long DNA Polymerase is a high performance, hot-start complex of enzymes and additives optimized for the sensitive DNA amplification of a wide range of challenging DNA templates including complex mammalian genomic DNA and long PCR amplicons greater than 20 Kb. The Hot-Start technology is based on a new-generation PCR buffer formulation in conjunction with a proprietary antibody-mediated chemistry. RadiantTM GC-Long DNA Polymerase exhibits significantly higher fidelity than wild-type Taq with an error rate of 5.0 x 10-5. The polymerase is ideal for multiplexing, low-copy assays, and the amplification of challenging GC-rich and AT-rich targets as well as long PCR of amplicons > 20Kb. RadiantTM GC-Long DNA Polymerase is engineered for robust, superior PCR and is supplied with a highly optimized, new-generation 5x buffer system which provides exceptional sensitivity and ease of use. Storage RadiantTM GC-Long 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 GC-Long DNA Polymerase 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 GC-Long Buffer: The 5x reaction buffer contains 15mM MgCl2, 5mM dNTPs, proprietary PCR enhancers and has been optimized for maximum efficiency, sensitivity and success with difficult amplicons. We do not suggest the use of additional PCR enhancers, dNTPs or MgCl2 . 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 55°C annealing temperature. 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. 15 seconds per kilobase(Kb) is recommended for amplification from eukaryotic genomic DNA or cDNA up to 5 Kb. For longer amplicons > 5Kb, we recommend between 40 seconds and 1 minute per Kb. Quality Control RadiantTM GC-Long DNA Polymerase is tested extensively for robust activity, processivity, efficiency, heat activation, sensitivity, absence of nuclease contamination and absence of nucleic acid contamination. RadiantTM GC-Long 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.