For Research Use Only.

Figure: Amplification of human GAPDH gene target with 2X HoTaq Real-time PCR Kit. Amplification curves are shown for ten fold dilutions of 0.0002pM to 20pM of plasmid. Inset shows the standard curve data.

Description:This is a high performance real-time PCR reagent. It utilizes MCLAB\'s proprietary quantitative PCR technology.Advantages:2x HoTaq PCR Reaction Mix products are superior in amplifying difficult templates comparing with similar products from other suppliers.- This is the amplification of GPIIB gene (70% G+C) .- 10 ~ 10K copies from 30pg human genomic DNA have been detected.Application:Probe based quantitative PCR: including DNA quantification, 2-step RT PCR, SNP analysis, etc.Primer and probe design: 1. To achieve the best performance, appropriate software, such as ABI Primer Express^TM, should be used to design primers with 50°C~65°C annealing temperature and 68°C~70°C for probes with 17~30 nucleotides in length2. Amplicon size should be small, <150bp3. Avoid secondary structures in primers and probes4. Avoid more than 3 consecutive Gs in primers and probes5. Primers should not have complementary 3\' -ends6. 17 ~ 30 nucleotides in lengthRecommended Reaction Conditions: :95°C, 10 minutes. -> (95°C, 5 seconds. -> 60°C, 30 seconds.) for 50 cycles.Recommended Storage Condition:-20°CNotes:To achieve accurate quantification, it is highly recommended to do replicates and to reduce pipetting errors.Reference: 1. Holland, P. M., Abramson, R. D., Watson, R., and Gelfand,2. D. H. 1991. Proceedings of the National Academy of Sciences USA 88:7276-7280.3. Livak, K. J., Flood, S. J. A., Marmaro, J., Giusti, W., and Deetz, K. 1995. PCR Methods and Applications 4:357-362.4. Lee, L. G., Connell, C. R., and Bloch, W. 1993 Nucleic AcidsResearch 21:3761-3766.