Maxima H Minus Reverse Transcriptase (200 U/µL)逆转录酶

货号：EP0751,EP0752,EP0753

规格：2000 units,10000 units,4 x 10000 units

价格：692,2182,6990

产品类型：反转录酶

品牌：Thermo Fisher

Thermo Scientific Maxima H Minus Reverse Transcriptase (RT) was developed throughin vitroevolution of M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity but lacks RNase H activity due to mutation in RNase H domain of M-MuLV RT. The engineered enzyme features dramatically improved thermostability, 50X higher processivity, robustness and increased synthesis rate compared to wild type M-MuLV RT.The eliminated RNase H activity enables the enzyme to produce very long RNA transcripts up to 20 kb. Due to its high thermostability, the enzyme maintains full activity during the entire reverse transcription reaction and generates high yields of cDNA. The reaction temperature can be increased up to 65°C for efficient transcription of RNA regions with a high secondary structure or to improve specificity using gene specific primers. The extremely high processivity of Maxima H Minus enzyme results in increased resistance to common reaction inhibitors, such as guanidine, formamide and ethanol.

Maxima 逆转录酶 (RT) 通过 M-MuLV RT 的体外进化制得。该酶具有 RNA 依赖性和 DNA 依赖性的聚合酶活性，但由于 M-MuLV RT 的 RNase H 域中的突变而缺乏 RNase H 活性。与野生型 M-MuLV RT 相比，该工程化酶具有显著提高的热稳定性，高50倍 的持续合成能力，稳健性和提升的合成速率。

消除 RNase H 活性使该酶能够产生长达 20 kb的 RNA 转录本。由于其热稳定性较高，该酶在整个逆转录反应过程中保持完整活性，并产生高产量的 cDNA。反应温度可提升至 65°C，以高效转录具有复杂二级结构的 RNA 可以进行高效的转录，或利用基因特异性的引物提高特异性。Maxima H Minus 酶具有极高的生产力，可提高对常见反应抑制剂（例如胍，甲酰胺和乙醇）的抗性。

产品优势

•热稳定性—在反应混合物中于 50°C 孵育60分钟后，具有 90% 的活性。

•最高活性温度 65°C

•全长 cDNA 的高产量，高达 20 kb

• 高敏感性—可从较宽范围的总 RNA 量（1 pg 至 5 µg）进行可重现的 cDNA 的合成

•高效—15 至 30 分钟内完成 cDNA 的合成

• 对常见反应抑制剂的抗性增强

•整合了修饰的核苷酸

应用

•用于 RT-PCR 和实时 RT-PCR 的第一链 cDNA 合成。

•在高温下进行逆转录，以减少二级结构的影响。

•用于克隆和表达的cDNA 的合成。

•用于微阵列的标记 cDNA 探针的产生。

•通过引物延伸分析 RNA。

注

•也可用：：Maxima H Minus 第一链 cDNA 合成试剂盒。

Highlights• Thermostable—90% active after incubation at 50°C for 60 minutes in a reaction mixture.• Active up to 65°C• High yields of full-length cDNA up to 20 kb• High sensitivity—reproducible cDNA synthesis from a wide range of starting total RNA amounts (1 pg to 5 µg)• Efficient—complete cDNA synthesis in 15 to 30 minutes• Increased resistance to common reaction inhibitors• Incorporates modified nucleotidesApplications• First strand cDNA synthesis for RT-PCR and real-time RT-PCR.• Reverse transcription at elevated temperatures to reduce effects of secondary structure.• Synthesis of cDNA for cloning and expression.• Generation of labeled cDNA probes for microarrays.• Analysis of RNA by primer extension.Note• Also available: :Maxima H Minus First Strand cDNA Synthesis Kit.Reduced effect of reaction inhibitorsEnzyme activity was determined in a standard activity assay using rection mixtures containing different concentrations of guanidinium chloride (0 to 750mM) or ethanol (0 to 30%).

技术参数

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