Radiant Hot-Start Taq Polymerase 4 x 50 µl, MgCl2, 5x HS Reaction Buffer €“ 8 x 1ml, Pack Size & Concentration: 4 x 250 units, 5u/µl

Radiant™ Hot-Start Taq DNA Polymerase is a highly purified, high performance Hot-Start DNA Polymerase 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.

Radiant™ Hot-Start Taq Polymerase exhibits 5´-3´ DNA polymerase activity with an error-rate of wild-type Taq ( 2.0 x 10-5). The polymerase is ideal for genotyping, colony PCR, multiplexing, low-copy assays, and the amplification of challenging targets susceptible to mispriming.

Radiant™ Hot Start Taq 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
Radiant™ Hot-Start Taq 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, Radiant™ Hot-Start Taq 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
Radiant™ 5x HS Taq Reaction Buffer: The 5x HS Taq 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.

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.