M-MLV Reverse Transcriptase: A Versatile Enzyme for cDNA Synthesis
M-MLV Reverse Transcriptase is an efficient and widely used enzyme for cDNA synthesis from RNA templates. It features high processivity and fidelity, making it suitable for a wide range of applications, including gene expression analysis, cloning, and sequencing.
M-MLV Reverse Transcriptase: An Enzyme for cDNA Synthesis
M-MLV Reverse Transcriptase (RTase) is an enzyme derived from the Moloney murine leukemia virus (M-MLV) that catalyzes the synthesis of complementary DNA (cDNA) from RNA templates. It is a versatile and widely used enzyme in molecular biology research due to its high processivity, fidelity, and ability to handle long RNA molecules.
Properties and Applications
M-MLV RTase possesses several key properties that make it a valuable tool for cDNA synthesis:
- High Processivity: The enzyme can efficiently elongate cDNA strands, synthesizing long stretches of DNA without pausing or detaching from the RNA template.
- High Fidelity: M-MLV RTase incorporates nucleotides accurately, minimizing errors during cDNA synthesis. This fidelity is crucial for maintaining the integrity of the genetic information being copied.
- Ability to Handle Long RNA Molecules: Unlike some other RTases, M-MLV RTase can effectively reverse transcribe long RNA molecules, making it suitable for cDNA synthesis from a wide range of RNA sources.
- Thermostability: M-MLV RTase is relatively stable at elevated temperatures, allowing for cDNA synthesis under mild conditions.
M-MLV RTase has a broad range of applications in molecular biology research, including:
- Gene Expression Analysis: cDNA synthesized using M-MLV RTase can be used to quantify gene expression levels using techniques such as real-time PCR or qPCR.
- Cloning: cDNA generated with M-MLV RTase can be inserted into cloning vectors for gene cloning and expression studies.
- Sequencing: cDNA derived from M-MLV RTase can be used for DNA sequencing to determine the nucleotide sequence of genes or transcripts.
- Synthesis of cDNA Libraries: M-MLV RTase is a key component in the construction of cDNA libraries, which represent the entire mRNA population of a cell or organism.
Advantages over Other RTases
M-MLV RTase offers several advantages over other RTases commonly used for cDNA synthesis:
- Higher Processivity: Compared to AMV RTase, M-MLV RTase can synthesize longer cDNA strands without disengaging from the RNA template.
- Reduced RNase H Activity: Unlike some other RTases, M-MLV RTase has minimal RNase H activity, which helps preserve the integrity of the RNA template during cDNA synthesis.
- Tolerance to Impurities: M-MLV RTase is relatively tolerant of impurities that may be present in RNA samples, making it less susceptible to inhibition.
- Availability of Modified Versions: Modified versions of M-MLV RTase, such as RNase H-deficient and thermostable variants, are available for specific applications.
M-MLV Reverse Transcriptase is a versatile and powerful enzyme that has become an essential tool in molecular biology research. Its high processivity, fidelity, ability to handle long RNA molecules, and relatively low RNase H activity make it a preferred choice for cDNA synthesis in a wide range of applications. With its well-established properties and extensive track record, M-MLV RTase is likely to continue playing a significant role in molecular biology research for years to come.
MMLV Reverse Transcriptase is an enzyme derived from the Moloney murine leukemia virus (M-MLV) that has been engineered through mutation screening to exhibit enhanced performance and stability. Unlike conventional M-MLV RTases, this modified version offers superior thermostability, enabling efficient reverse transcription at elevated temperatures of 50-60°C. This high-temperature tolerance effectively eliminates the adverse effects of RNA secondary structures and non-specific factors on cDNA synthesis, resulting in more robust and reliable cDNA generation.
- High Thermostability: Performs reverse transcription at high temperatures (50-60°C), ensuring stable reaction conditions.
- Enhanced Reverse Transcription Efficiency: Guarantees efficient cDNA synthesis and high success rates, even with low-concentration RNA templates.
- First-strand cDNA synthesis for RT-PCR and RT-qPCR: Generates high-quality cDNA from RNA templates for downstream applications such as real-time PCR and quantitative PCR.
- Synthesis of cDNA for cloning and expression studies: Produces cDNA suitable for gene cloning and expression studies, enabling the study of gene function and protein production.
SDS-PAGE electrophoresis analysis demonstrates the purity and integrity of MMLV Reverse Transcriptase compared to similar products. The enzyme exhibits a single distinct band at the expected molecular weight, indicating its high purity and lack of contaminating proteins.
Comparative testing with competitive enzymes
Accelerated thermal stability
|Product Name||M-MLV Reverse Transcriptase|
|Source||Recombinant expression in Escherichia coli|
|Enzyme Activity||200 U/µL|
|Storage Conditions||-20 ±5°C|
|Molecular Weight||74 kDa|
|Storage Buffer||20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.01% NP-40, 50% Glycerol|
|Activity Definition||1 unit (U) is the amount required to incorporate 1 nmol of dTTP into acid-insoluble material in a 50 μL reaction system with Poly(rA)•Oligo(dT)15 as the template at 37°C for 10 minutes.|
|Quality Control||No detectable endonuclease, DNase, and RNase activities. PCR testing shows no residual host genomic DNA.|
|Shelf Life||24 months|
Elevate your reverse transcription with M-MLV Reverse Transcriptase, a meticulously engineered enzyme that not only reduces RNase H activity but also enhances thermal stability. Compared to wild-type M-MLV, this variant can synthesize the first strand cDNA at elevated temperatures, showcasing its versatility and adaptability. It operates with normal activity at 50-60°C, offering increased specificity, higher yield, and the remarkable ability to synthesize cDNA up to 12 kb in length.
|Component No.||Component Name||Specifications|
|1||5× First-Strand Buffer||0.4 mL, 2 mL, 4 mL|
|2||M-MLV Reverse Transcriptase||20000 U, 100000 U, 200000 U|