M-MLV 反转录酶 (200 U/μL)

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Invitrogen™

M-MLV 反转录酶 (200 U/μL)

Name: M-MLV 反转录酶 (200 U/μL)
SKU: 28025013
Price: 3100.00 CNY

M-MLV 反转录酶是一种以单链 RNA、DNA 或 RNA:DNA 杂交体为模板合成互补 DNA 链的重组 DNA了解更多信息

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货号	反应次数
28025013	200 次反应
28025021	1,000 次反应

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货号 28025013

价格（CNY)

3,100.00

Each

添加至购物车

反应次数:

200 次反应

请求批量或定制报价

价格（CNY)

3,100.00

Each

添加至购物车

M-MLV 反转录酶是一种以单链 RNA、DNA 或 RNA:DNA 杂交体为模板合成互补 DNA 链的重组 DNA 聚合酶。与 AMV RT 相比，莫洛尼鼠白血病病毒反转录酶 (M-MLV RT) 缺乏 DNA 核酸内切酶活性，RNase H 活性较低。该酶的特点：

•热稳定性—在 37°C 下活性极佳
•cDNA 大小—M-MLV 可用于合成长达 7 kb 的第一链 cDNA
•应用—合成第一链 cDNA、引物延伸、dsDNA 测序、cDNA 文库和 RT-PCR

来源
纯化自表达质粒所带 M-MLV pol 基因的大肠杆菌

性能和质量检测
SDS-PAGE 纯度；脱氧核糖核酸内切酶、脱氧核糖核酸外切酶及核糖核酸酶检测；cDNA 产物的得率和长度

单位定义
一个单位的 M-MLV RT 是指在 37°C、10 分钟条件下使用 poly(A)oligo(dT)₂₅ 作为模板-引物将 1 nmole 脱氧核糖核酸掺入酸性可沉淀材料所需的酶量。

单位反应条件
50 mM Tris-HCl（pH 值 8.3）、40 mM KCl、6 mM MgCl₂、1 mM DTT、0.5 mM [³H]dTTP、0.1 mM poly(A)、0.1 mM oligo(dT)₂₅、0.1 mg/mL BSA 和酶，50 µL，10 min，37°C。

仅供科研使用。不可用于诊断程序。

规格

最终产品类型第一链 cDNA

产品规格管装

反应次数200 次反应

最佳反应温度37°C

数量200 次反应；40,000 单位

反应形式单独组分

试剂类型反转录

逆转录酶M-MLV

核糖核酸酶 H 活性是

运输条件湿冰

尺寸（最终产品）长达 7 kb

原始材料RNA

技术反转录

最大浓度200 U/μL

反应速度50 min。

Unit SizeEach

内容与储存

• M-mLV RT，200 μL (200 U/μL)
• 5X 第一链缓冲液，1 mL
• DTT，500 μL (100 mM)

储存在 –20°C 下。

常见问题解答 (FAQ)

PCR反应要加入多少第一链cDNA反应产物？

尽管实际体积取决于用于第一链合成反应的RNA起始量和靶标基因的丰度，但我们建议使用10%的第一链cDNA反应产物进行PCR反应。

How much of the first-strand cDNA reaction should I load for PCR?

While the volume is dependent on the starting amount of RNA used for the first-strand synthesis and the abundance of the target gene, we'd recommend starting with 10% of the first-strand reaction for your PCR reaction.

How can reverse transcriptases be inactivated?

The enzymes can be inactivated by adding a chelating agent such as EDTA. Alternatively, with the exception of ThermoScript RT and Thermo-X RT, the enzymes can be heat inactivated at 70 degrees C for 10 min.

ThermoScript RT should be heated to 85 degrees C for 5 min for complete inactivation.

For Thermo-X RT, if using an oligo(dT) primer, add EDTA to the reaction at a final concentration of 5 mM. Inactivate the reaction by heating at 90 degrees C for 5 min.

What is the highest temperature that SuperScript III , SuperScript II, MMLV, or ThermoScript can be used?

The optimal temperature for SuperScript III RT is 50 degrees C, and can be used up to 55 degrees C. For some qRT-PCR reactions where gene-specific primers are used, you can do the RT reaction at 60 degrees C. The optimal temperature for SuperScript II RT is 42 degrees C, and can be used up to 50 degrees C. Optimal temperature for MMLV is 42 degrees C. ThermoScript RT shows optimal activity at 60 degrees C, and can be used at temperatures as high as 70 degrees C (for amplicons expected to be 1 kb or less). For PCR products expected to be greater than 1 kb, a maximum first strand synthesis temperature of 60-65 degrees C is suggested. Be sure your first-strand primer anneals at the high temperature, especially when gene-specific primers are used for high-temperature stable reverse transcriptases. We recommend oligo (dT)20 for cDNA synthesis when using an oligo (dT) primer for first-strand synthesis with these enzymes.

Can I use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase (Cat. No. 28025013, 28025021)? Can other reverse transcriptases, such as SuperScript reverse transcriptase, be used in the same way?

Yes, you can use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase.

We have not tested this for SuperScript reverse transcriptases, so we cannot guarantee it would also work with those products.

This article can be used as a reference for additional information.

Find additional tips, troubleshooting help, and resources within our Reverse Transcription and RACE Support Center.

View all M-MLV 反转录酶 (200 U/μL) FAQs

引用和文献 (4)

引用和文献

Abstract

Raloxifene Upregulated Mesangial Cell MMP-2 Activity via ER-ß Through Transcriptional Regulation.

Authors:Fang M, Wu XC, Huang W,

Journal:Cell Biochem Biophys

PubMed ID:23471663

'Raloxifene, a second-generation selective estrogen receptor modulator, exerts estrogen-like effects in specific tissues. In this present study, we examined the effect of raloxifene on mesangial cell matrix metalloproteinase-2 (MMP-2) activity in streptozotocin-induced diabetic mice. Raloxifene increased the MMP-2 level in a dose-dependent and receptor-mediated manner. An antibody against estrogen receptor-ß ... More

Single cell rt-PCR identification of odorant receptors expressed by olfactory neurons.

Authors:Malnic B,

Journal:Methods Mol Biol

PubMed ID:23585038

'Mammals have between 400 and 1,300 functional odorant receptor (OR) genes in their genomes. Each olfactory sensory neuron in the nose expresses only one single type of OR out of this vast repertoire. The OR expressed by an olfactory sensory neuron determines its functional activity and wiring to the olfactory ... More

Fast-mode duplex qPCR for BCR-ABL1 molecular monitoring: innovation, automation, and harmonization.

Authors:Gerrard G, Mudge K, Foskett P, Stevens D, Alikian M, White HE, Cross NC, Apperley J, Foroni L,

Journal:Am J Hematol

PubMed ID:22566190

Reverse transcription quantitative polymerase chain reaction (RTqPCR)is currently the most sensitive tool available for the routine monitoring of disease level in patients undergoing treatment for BCRABL1 associated malignancies. Considerable effort has been invested at both the local and international levels to standardise the methodology and reporting criteria used to assess ... More

Transforming growth factor-ß is required for vasculogenic mimicry formation in glioma cell line U251MG.

Authors:Ling G, Wang S, Song Z, Sun X, Liu Y, Jiang X, Cai Y, Du M, Ke Y,

Journal:Cancer Biol Ther

PubMed ID:22104964

Both vasculogenic mimicry (VM) and transforming growth factor-ß (TGFß) are positively correlated with malignancy in glioma. Accordingly, we supposed that TGFß might be related with VM, and aimed to detect whether TGFß could influence VM formation in two glioma cell lines U251MG and SHG44, which were different in malignancy. We ... More