MasterAmp™ Tth DNA Polymerase/MasterAmp™ Tth DNA聚合酶
货号：TTH72250
规格：250 U 5 U/µL
价格：2331
产品类型：PCR 及DNA聚合酶
品牌：Lucigen
| MasterAmp™ Tth DNA Polymerase is a thermostable DNA Polymerase from Thermus thermophilus. The enzyme displays optimal activity at temperatures between 70°C and 74°C, and it is useful for performing the polymerase chain reaction (PCR)* or other reactions requiring DNA synthetic activity at high temperatures. Tth DNA polymerase has an intrinsic 5′→3′ exonuclease activity, but lacks a 3′→5′ (proofreading) nuclease activity. Furthermore, Tth has a potent Mn-dependent reverse transcriptase activity.The enzyme is provided with MasterAmp Tth 20X PCR Buffer, MasterAmp 10X PCR Enhancer (with betaine)+, and 25 mM MgCl2, and MnSO4 Solutions to allow optimization of individual template-primer pair combinations. MasterAmp™Tth DNA聚合酶是Thermus thermophilus的热稳定DNA聚合酶。该酶在70°C至74°C的温度下显示最佳活性，可用于在高温下进行聚合酶链反应（PCR）*或其他需要DNA合成活性的反应。Tth DNA聚合酶具有固有的5'→3'核酸外切酶活性，但缺乏3'→5'（校对）核酸酶活性。此外，Tth具有强大的Mn依赖性逆转录酶活性。该酶配有MasterAmp Tth 20X PCR缓冲液，MasterAmp 10X PCR增强剂（含甜菜碱）+，25 mM MgCl2和MnSO4溶液，可优化单个模板引物对组合。The presence of betaine (trimethyl glycine) in the MasterAmp 10X 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 Betaine, a natural osmoprotectant, lowers the melting temperature of G+C rich regions to a temperature more similar to A+T rich regions.This results in destabilization of double-stranded DNA which limits polymerase pausing, thereby increasing the yield of full-length product. 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. 甜菜碱（三甲基甘氨酸）在MasterAmp 10X PCR增强剂中的存在可显着提高许多靶序列的产量和扩增特异性，尤其是那些含有高G + C含量或二级结构甜菜碱（天然的渗透保护剂）的序列，可降低甜菜碱的解链温度。G + C富集区的温度与A + T富集区的温度更相似，这会导致双链DNA不稳定，从而限制了聚合酶的暂停，从而提高了全长产物的产量。此外，甜菜碱还可以通过保护DNA聚合酶免受热变性而增强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目标序列而变化。 这种来自嗜热栖热菌的重组酶在高达~95℃时仍具有DNA聚合酶活性，以及逆转录酶活性。高反应温度可以减少非特异性扩增和模板二级结构。随酶附带MasterAmp PCR Enhancer。 |
| 优点： |
| ▪ Flexible: Use a single enzyme for both PCR and RT-PCR applications, greatly simplifying reaction optimization and setup▪ Thermostable: Enables amplification of DNA and RNA targets containing high degrees of secondary structure▪ 灵活性：在PCR和RT-PCR应用中都使用单一酶，大大简化了反应的优化和设置▪ 热稳定性：能够扩增包含高度二级结构的DNA和RNA靶标 |
| 数据： |
| Enzyme Characteristics:5′→3′ Exonuclease Activity ...................Yes 3′→5′Exonuclease Activity ...............................No Terminal Transferase Activity ..................Yes Extension Rate (72°C) ..............................2-4 kb/minProcessivity ...............................................50-60 basesHalf-life .............................................10 min (97°C), 40 min (95°C) Reverse Transcriptase Activity (Mn2+ dependent) .......... Yes |
| 组成成分： |
| ▪ MasterAmp™ Tth DNA Polymerase (20 Unit/µL)：储存在-20°C▪MasterAmp™ 10X PCR Enhancer (with Betaine)：储存在-20°C▪ MgCl2, 25 mM：储存在-20°C▪MnSO4, 25 mM：储存在-20°C▪ 20X Tth DNA Polymerase Reaction Buffer：储存在-20°C |
| 相关产品： |
| ▪ 2.5 mM dNTP Mix, PCR Grade（# 30030）▪ 2'-dNTP (All 4 Mixed)（# D08104） |
| 参考文献： |
| 1. Myers, T.W. and Gelfand, D.H. (1991) Biochem. 30, 7661. 2. Mytelka, D.S. and Chamberlin, M.J. (1996) Nucl. Acids Res. 24, 2774.3. Henke, W. et al., (1997) Nucl. Acids Res. 25, 3957.4. Weissensteiner, T. and Lanchbury, J.S. (1996) BioTechniques 21, 1102. 5. Rees, W.A. et al., (1992) Biochemistry 32, 137. |
|
技术参数 产品优点 - Flexible: Use a single enzyme for both PCR and RT-PCR applications, greatly simplifying reaction optimization and setup
- Thermostable: Enables amplification of DNA and RNA targets containing high degrees of secondary structure
- 灵活性：在PCR和RT-PCR应用中都使用单一酶，大大简化了反应的优化和设置
- 热稳定性：能够扩增包含高度二级结构的DNA和RNA靶标
产品应用 - DNA的PCR扩增
- 改进了针对具有高度二级结构的DNA模板的PCR扩增
- RNA的一步式RT-PCR
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