Reverse transcriptases are enzymes encoded in retroviruses viral genome. The enzyme is responsible for transcription of the viral RNA to produce a dsDNA that can be inserted into the host genome.
Reverse transcriptases are multifunctional enzymes. These enzymes exhibit an RNA and DNA directed polymerase activity. In addition reverse transcriptases catalyze the degradation of RNA in an RNA-DNA hybrid. The exonucleolytic activity proceeds in a 5' ---> 3' direction. The RNA or DNA directed activity requires a template (RNA or DNA) and a primer. The following is a schematic illustration of the reaction:

Unit definition: One unit incorporates 1 nanomole of tritiated dTMP into acid insoluble productsusing poly(A)•oligo(dT) 12-18 as the template-primer in 20 minutes at 37° C.

Applications
HIV reverse transcriptase is used for research on the AIDS primer. However it can be substituted for AMV reverse transcriptase, which is mainly used to transcribe mRNA into double stranded cDNA, that can be inserted into prokaryotic vectors. The enzyme can also be used with either single stranded DNA or RNA templates to make probes for use in hybridization experiments. It can be used for labeling the termini of DNA fragments with protruding 5' termini. The enzyme can also be used to sequence DNAs by the dideoxy chain termination method of Sanger when the Klenow fragment of E. coli DNA polymerase I, or the T7 DNA polymerase yield unsatisfactory results.

Reagents
0.05 M Tris, pH 8.3, containing 0.008 M MgCl2
1 mg/ml polyadenylic acid in water (poly A)
DNA primer:Oligo d(T)12-18
1 μ mole dTTP/mL stock solution
[methyl-3H]-Thymidine 5'-triphosphate (3H-dTTP)
dTTP-3H-dTTP working mix: Add 1-2 μL 3H-dTTP per mL of 100 nmol/mL dTTP in order to obtain 1 to 1.5 x 105 cpm/mL
1% bovine serum albumin
10% perchloric acid
1% perchloric acid
Buffer substrate reaction mixture: Prepare fresh, immediately before use:
For each 1mL of reaction mixture required mix:
0.7 mL Tris/HCl, pH 8.3, 0.008M MgCl2
0.3 mL 1 mg/mL poly(A) RNA template
0.005 mL 0.02 mg/mL oligo d(T)12-18 DNA primer
0.02mL 1% BSA
Enzyme
dilute as needed wtih 0.05M Tris/HCl, pH 8.3, 0.008M MgCl2 containing 0.1 mg/mL (1%) BSA
Procedure
Pipette into each tube as follows:
Buffer substrate mix：0.1 mL
dTTP-3H3-dTTP：0.1 mL
Enzyme：5-10 μL
Incubate 20 minutes at 37° C. Stop reaction by adding 1 ml 10% cold perchloric acid. Filter through 0.2μ manifold filters used with Millipore vacuum manifold. Wash four times using 2mL 1% cold perchloric acid/wash. Transfer filter to scintillation vials. Add 2mL Cellosolve (or 2-methoxyethanol) to dissolve filter. Filters become opaque upon addition of Cellosolve. Make sure filters are dissolved before proceeding. Add 10mL scintillation cocktail and count.
Calculation

规格

产品名称

重组HIV逆转录酶

英文别名

规格或纯度

生物活性, 重组, 无DNA酶和RNA酶, EnzymoPure™, 大肠杆菌表达，≥5000 U/mg protein; 蛋白浓度见COA

生物活性

≥5000 U/mg protein

表达系统

E. coli

种属

HIV

来源

重组表达

CAS编号和信息

9068-38-6

酶学委员会编号

2.7.7.49

分子类型

酶

储存与运输

物理形态

液体

储存缓冲液

10mM Potassium phosphate, 1mM DTT, 20% Glycerol, pH 7.4.

浓度

大肠杆菌表达，≥5000 U/mg protein; 蛋白浓度见COA

储存条件

-20°C储存

运输条件

超低温运输

单位定义

在 37℃、pH 8.3 条件下，以 poly (A)/oligo (dT)12-18为模板 / 引物，20 分钟内将 1 纳摩尔氚标记的 d-TMP 掺入酸可沉淀产物中所需的酶量。

高级数据

质检证书(CoA,COO,BSE/TSE 和分析图谱)

C of A & Other Certificates(BSE/TSE, COO):

输入批号以搜索分析图谱:

通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！

找到4个结果

批号(Lot Number)	证书类型	货号
I2524662	分析证书	R128560
C2426339	分析证书	R128560
I1818102	分析证书	R128560
I1818101	分析证书	R128560

技术文档和文章

蛋白样品制备流程

目录：技术文章

探索主题:

蛋白样品制备

Protein sample preparation process

目录：技术文章

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Protein sample preparation

分子生物学实验中常用聚合酶的类型、特性与应用

目录：技术文章

探索主题:

PCR DNA聚合酶等温扩增 qPCR RNA聚合酶聚合酶反转录酶高保真扩增体外转录

Types, Properties, and Applications of Commonly Used Polymerases in Molecular Biology Experiments

目录：技术文章

探索主题:

PCR qPCR

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

浓度

体积

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

体积 1 (V1)

浓度 2 (C2)

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

质量（小瓶）

所需的复溶浓度

需要帮助选择级别？

我们的级别选择指南涵盖了目录中所有的变体的纯度、稳定剂状态和应用适用性。

查看 Bioactive 级别指南 → 查看 Recombinant 级别指南 → 查看 DNase, RNase free 级别指南 → 查看 EnzymoPure™ 级别指南 →

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重组HIV逆转录酶

概述

规格

高级数据

摩尔浓度计算器

稀释计算器

复溶计算器

需要帮助选择级别？

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