| 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. FailSafe™PCR PreMix选择试剂盒包含独特的热稳定DNA聚合酶混合物和一组十二种反应PreMixes。 这套全面的试剂专为满足每种PCR需求而设计。 任何模板：例程； 困难（例如高GC含量或二级结构）; 或长（大约20 kb的长度），可以使用FailSafe PCR轻松扩增。 而且由于FailSafe PCR酶混合物的保真度比单独Taq DNA聚合酶高至少三倍，因此PCR产品适用于下游应用，例如克隆，测序，表达和突变分析。 FailSafe PCR酶混合物是一种酶混合物，其中包含3'→5'校对酶，以确保高保真度。 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. 12个FailSafe PCR 2X PreMix包含具有所有4种dNTP的缓冲盐溶液，各种量的MgCl2和FailSafe PCR Enhancer（含甜菜碱），用户只需向每个PreMix中添加模板，引物和FailSafe PCR酶混合物即可，并放大。 结果将在凝胶上清楚显示哪种PreMix最适合该模板/引物对组合。FailSafe PCR增强剂中甜菜碱（三甲基甘氨酸）的存在大大提高了许多靶序列的扩增效率和特异性，尤其是那些含有高G + C含量或二级结构的靶序列。FailSafe PCR增强剂中甜菜碱（三甲基甘氨酸）的存在大大提高了许多靶序列的扩增效率和特异性，尤其是那些含有高G + C含量或二级结构的靶序列。此外，甜菜碱还可以通过保护DNA聚合酶免于热变性来增强PCR。甜菜碱的作用似乎与所用的聚合酶无关，尽管扩增所需的甜菜碱浓度随靶序列而变化。 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. 一旦确定了最佳的PreMix，即可使用针对模板/引物对组合定制的带有PreMix Choice的FailSafe™PCR系统实现一致的扩增。从与您选择的FailSafe PCR PreMixes配对的不同规格的FailSafe PCR酶混合物中进行选择。 单独的FailSafe™PCR 2X PreMixes和仅FailSafe™酶混合物也可单独购买 FailSafe Enzyme Mix是一种独特的热稳定DNA聚合酶和3’→5’校正外切核酸酶的混合物，能够扩增最困难的模板。FailSafe酶提供的保真度比Taq DNA聚合酶高3倍。其扩增PCR产物能力可达20kb。FailSafe Enzyme Mix的错配率约为27,000-30,000分之一。 |
| 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)▪ PCR和多重PCR▪ 困难模板PCR▪ 高灵敏度PCR▪ PCR扩增长达20kb的任何序列 |
| 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.图1.使用FailSafe™PCR PreMix选择试剂盒扩增富含GC的模板。 扩增了人类ApoE基因75％的富含GC的区域。 M，分子标记； 泳道1，使用MasterAmp Taq DNA聚合酶的标准缓冲液条件； 泳道A-L，分别用12个FailSafe PCR 2X PreMixes进行扩增。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.图2.使用FailSafe™PCR PreMix K进行一致的扩增。使用分子标记FailSafe PCR 2X PreMix K.M进行了图1的ApoE基因的十二次重复扩增反应.Primerquest或Primer 3可简化引物设计以消除可能的引物二聚体或交叉反应性。 |
| 相关产品： |
| ▪ FailSafe™ PCR System(#FS9901K/FS99100/FS99250)▪ 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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