| The FailSafe™ PCR PreMix Selection Kit contains a unique blend of thermostable DNA polymerases and a set of twelve reaction PreMixes. This comprehensive set of reagents was specifically designed to meet every PCR need. Any template: routine; difficult (e.g., high GC content or secondary structure); or long (approximately 20 kb in length), can easily be amplified with FailSafe PCR. And because the FailSafe PCR Enzyme Mix provides fidelity at least three times higher than Taq DNA polymerase alone, PCR products are suitable for downstream applications such as cloning, sequencing, expression, and mutation analysis. The FailSafe PCR Enzyme Mix is an enzyme blend containing a 3′→5′ proofreading enzyme for high fidelity. The 12 FailSafe PCR 2X PreMixes contain a buffered salt solution with all 4 dNTPs, and various amounts of MgCl2, and FailSafe PCR Enhancer (with betaine).The user simply adds template, primers, and the FailSafe PCR Enzyme Mix to each of the PreMixes and amplifies. The results will clearly show, on a gel, which PreMix is best for that template/primer pair combination. The presence of betaine (trimethyl glycine) in the FailSafe PCR Enhancer substantially improves the yield and specificity of amplification of many target sequences, especially those containing a high G+C content or secondary structure.In addition, betaine also may enhance PCR by protecting DNA polymerases from thermal denaturation.The effects of betaine seem to be independent of the polymerase used, though the concentration of betaine required for amplification varies with the target sequence.Once the optimal PreMix has been determined, consistent amplification is achieved with the FailSafe™ PCR System with PreMix Choice customized to your template/primer pair combination. Choose from different sizes of FailSafe PCR Enzyme Mix paired with the FailSafe PCR PreMixes of your choice. Individual FailSafe™ PCR 2X PreMixes and FailSafe™ Enzyme Mix Only are also available separately. |
| 优点： |
| ▪ Successful PCR, the first time and every time▪ PCR amplification of difficult or high-GC templates ▪ Multiplex PCR compatible ▪ Robust amplification of targets up to 20 kb long▪ High accuracy with 3-fold lower error rate than Taq DNA polymerase (error rate = 1 in 30,000)▪ Extremely high sensitivity and specificity using the PCR Enhancer Technology (with Betaine) |
| 数据： |
| Figure 1. Amplification of a GC-rich template using the FailSafe™ PCR PreMix Selection Kit. A 75% GC-rich region of the human ApoE gene was amplified. M, molecular marker; Lane 1, standard buffer conditions using MasterAmp Taq DNA polymerase; Lanes A-L, amplification with each of the 12 FailSafe PCR 2X PreMixes.Figure 2. Consistent amplification with FailSafe™ PCR PreMix K. Twelve repeat amplification reactions of the ApoE gene from Fig. 1 were performed using FailSafe PCR 2X PreMix K. M, molecular marker.Primerquest or Primer 3, may simplify primer design to eliminate possible primerdimers or cross-reactivity |
| 相关产品： |
| ▪ FailSafe™ Enzyme Mix Only(# FSE51100/FSE5101K) ▪ FailSafe™ PCR PreMix Selection Kit（# FS99060） ▪ FailSafe™ PCR 2X PreMix A(#FSP995A)▪ FailSafe™ PCR 2X PreMix B (#FSP995B)▪ FailSafe™ PCR 2X PreMix C (# FSP995C)▪ FailSafe™ PCR 2X PreMix D (# FSP995D)▪ FailSafe™ PCR 2X PreMix E (# FSP995E)▪ FailSafe™ PCR 2X PreMix F (# FSP995F)▪ FailSafe™ PCR 2X PreMix G (# FSP995G)▪ FailSafe™ PCR 2X PreMix H (# FSP995H )▪ FailSafe™ PCR 2X PreMix I (# FSP995I)▪ FailSafe™ PCR 2X PreMix J (# FSP995J)▪ FailSafe™ PCR 2X PreMixK (# FSP995K) ▪ FailSafe™ PCR 2X PreMix L (# FSP995L) |
| 参考文献： |
| 1. Mytelka, D.S. and Chamberlin, M.J. (1996) Nucl. Acids Res. 24, 2774. 2. Henke, W. et al., (1997) Nucl. Acids Res. 25, 3957. 3. Weissensteiner, T. and Lanchbury, J.S. (1996) BioTechniques 21, 1102. 4. Santoro, M.M. et al., (1992) Biochemistry 31, 5278.5. Roux, K.H. (1995) in: PCR Primer, Dieffenbach, C.W. and Dveksler, G.S. (eds.), CSH Press, New York, 53. |
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