Radiant™ 2X Hot-Start Taq Mastermix Description RadiantTM 2x Hot-Start Taq Mastermix is a ready-to-use, robust PCR mix containing our highly purified Hot-Start Taq DNA Polymerase, reaction buffer, ultra-pure dNTPs, MgCl2 and PCR enhancers. RadiantTM 2x Hot-Start Taq Mastermix is optimized for the sensitive DNA amplification of a wide range of DNA templates including complex mammalian genomic DNA and crude samples. The Hot-Start technology is based on a new-generation PCR buffer formulation in conjunction with a proprietary antibody-mediated chemistry. RadiantTM Hot-Start Taq Mastermix exhibits 5´-3´ DNA polymerase activity with an error-rate of wild-type Taq ( 2.0 x 10-5). The polymerase mix is ideal for genotyping, colony PCR, multiplexing, low-copy assays, and the amplification of challenging targets susceptible to mispriming. RadiantTM Hot Start Taq Mastermix is engineered for robust, superior PCR and is supplied as a ready-to-use 2x mix which provides exceptional sensitivity and ease of use. Storage RadiantTM 2x Hot-Start Taq Mastermix 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 2x Hot-Start Taq Mastermix is stable for 12 months from date of receipt. The Kit may also be stored at 4°C for 1 month. Important Considerations RadiantTM 2x Hot-Start Taq Mastermix: The 2x Mastermix is comprised of RadiantTM Hot-Start Taq DNA Polymerase, 2mM dNTPs, 6mM MgCl2, and PCR enhancers for maximum efficiency, sensitivity and success with difficult amplicons. We do not suggest the use of additional PCR 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 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. For shorter amplicons, a 1 second extension is sufficient. For Multiplex PCR, we suggest an initial annealing temperature gradient from 55°C to 65°C in order to determine the highest level of specificity. In addition, we recommend an initial extension time of 90 seconds and greater to maximize yield and specificity. Quality Control RadiantTM 2x Hot-Start Taq Mastermix is tested extensively for robust activity, processivity, efficiency, heat activation, sensitivity, absence of nuclease contamination and absence of nucleic acid contamination. RadiantTM 2x Hot-Start Taq Mastermix 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.