名称:反转录酶 III,SuperScript™ III Reverse Transcriptase 产地:invitrogen  货号: 18080-044 规格:10000u

描述
SuperScript® III 反转录酶 (RT) (SuperScript® III Reverse Transcriptase (RT)) 是 SuperScript® II RT 的专有突变体，在 50°C 具有活性，半衰期为 220 分钟，使用基因特异性引物 (GSP) 提高特异性，在所有 RT 中 cDNA 产量最高。 如果要进行特定基因的 RT-PCR 或从总 RNA 或 poly (A)+ RNA 样品生成 cDNA，该产品十分理想。 和 SuperScript® II 一样，它可以从单链 RNA、DNA 或 RNA:DNA 杂合体合成互补 DNA 链。 通过引入点突变对 SuperScript® III RT 进行遗传改造，这些点突变可以增加半衰期、降低 RNase 活性并增加热稳定性。 进行结构修饰后：
50°C 时半衰期为 220 分钟，获得最高的 cDNA 产量（图 1）
RNase H 活性降低，获得更多全长 cDNA
在 50°C 有最大活性，用 GSP 增加特异性
能增加 RT 单位而不抑制后续的 PCR

应用： 第一链 cDNA 合成、阵列标记、cDNA 文库、RT-PCR、引物延伸和 3´ 和 5´ RACE。

来源： 纯化自表达 M-MLV 的 pol 基因的大肠杆菌 (1,2)，通过突变增加了热稳定性和半衰期，降低了 RNase H 活性。

性能和质量测试： SDS-PAGE 纯度；脱氧核糖核酸内切酶、脱氧核糖核酸外切酶和核糖核酸酶分析（图 2）及 cDNA 产物的产量和长度。

单位定义： 一个单位的 SuperScript® III RT 是在 37°C 的反应体系中，使用 poly(A)•oligo(dT)12-18 作为模板•引物，在 10 分钟内将 1 纳摩尔脱氧核糖核苷酸掺入酸沉淀材料所需的酶量 (3)。

单位反应条件： 50 mM Tris-HCl (pH 8.3)、40 mM KCl、6 mM MgCl 2、1 mM DTT。 0.5 查看更多mM [ 3 H]dTTP、0.1 mM poly(A)、0.1 mM oligo(dT)12-18、0.1 mg/ml BSA 和酶，共 50 µl，温度 37°C，时间 10 分钟。

Description:

    SuperScript™ III Reverse Transcriptase (RT) is a proprietary mutant of SuperScript™ II RT that is active at 50°C and has a half-life of 220 minutes, providing increased specificity with Gene-Specific Primers (GSPs) and the highest     View More

    SuperScript™ III Reverse Transcriptase (RT) is a proprietary mutant of SuperScript™ II RT that is active at 50°C and has a half-life of 220 minutes, providing increased specificity with Gene-Specific Primers (GSPs) and the highest cDNA yield of all RTs. It is ideal for RT-PCR of a specific gene or generating cDNA from total or poly (A)+ RNA sample. Like SuperScript™ II, it synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. SuperScript™ III RT is genetically engineered by the introduction of point mutations that increase half-life, reduce RNase activity, and increase thermal stability. The structural modifications provide:

Half life of 220 minutes at 50°C for the highest cDNA yields (Figure 1)
Reduced RNase H activity for more full-length cDNA
Full activity at 50°C for increased specificity with GSP
Ability to increase RT units without inhibiting subsequent PCR
Applications: Synthesis of first-strand cDNA, array labeling, cDNA libraries, RT-PCR, primer extension, and 3´ and 5´ RACE.

Source: Purified from E. coli expressing the pol gene of M-MLV (1,2), mutagenized to increase thermal stability and half life and reduce RNase H activity.

Performance and Quality Testing: SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays (Figure 2) and yield and length of cDNA product.

Unit Definition: One unit of SuperScript™ III RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min. at 37°C using poly(A)•oligo(dT)12-18 as a template•primer (3).

Unit Reaction Conditions: 50 mM Tris-HCl (pH 8.3), 40 mM KCl, 6 mM MgCl 2 , 1 mM DTT. 0.5 mM [ 3 H]dTTP, 0.1 mM poly(A), 0.1 mM oligo(dT)12-18 , 0.1 mg/ml BSA, and enzyme in 50 µl for 10 min. at 37°C.

Contents and Storage:
SuperScript™ III RT is supplied with a vial of 5X first-strand buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2], and a vial of 100 mM DTT. Store at -20°C. Guaranteed stable for 6 months when properly stored

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