M-MLV Reverse Transcriptase (200 U/μL)
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M-MLV Reverse Transcriptase (200 U/μL)
Invitrogen™

M-MLV Reverse Transcriptase (200 U/μL)

M-MLV Reverse Transcriptase is a recombinant DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or anRead more
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Catalog NumberNo. of Reactions
28025013200 Reactions
280250211,000 Reactions
Catalog number 28025013
Price (USD)
406.00
Each
Add to cart
No. of Reactions:
200 Reactions
Request bulk or custom format
Price (USD)
406.00
Each
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M-MLV Reverse Transcriptase is a recombinant DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. Compared to AMV RT, Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) lacks DNA endonuclease activity and has a lower RNase H activity.

One unit of M-MLV 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)25 as template-primer.

Features of M-MLV Reverse Transcriptase include:
• Excellent thermostability with optimal activity at 37°C
• Can be used to synthesize first-strand cDNA up to 7 kb
• Multiple applications including synthesis of first-strand cDNA, primer extension, sequencing dsDNA, cDNA libraries, and RT-PCR

Source
Purified from E. coli expressing the pol gene of M-MLV on a plasmid

Performance and quality testing
SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product

Unit reaction conditions
50 mM Tris-HCl (pH 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, and enzyme in 50 μL for 10 min at 37°C

For Research Use Only. Not for use in diagnostic procedures.
Specifications
FormatStand-alone Enzyme
Reaction Speed50 min.
TechniqueReverse Transcription
Optimal Reaction Temperature37°C
Reverse TranscriptaseM-MLV
Ribonuclease H ActivityYes
Shipping ConditionWet Ice
Concentration200 U/μL
Final Product TypeFirst-Strand cDNA
No. of Reactions200 Reactions
Reaction FormatSeparate components
Reagent TypeReverse Transcription
Size (Final Product)Up to 7 kb
Starting MaterialRNA
Unit SizeEach
Contents & Storage

• M-mLV RT, 200 μL (200 U/μL)
• 5X First Strand Buffer, 1 mL
• DTT, 500 μL (100 mM)

Store at –20°C.

Frequently asked questions (FAQs)

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.

Can I use elevated temperatures for reverse transcription with Moloney Murine Leukemia Virus (M-MLV)?

When using GeneAmp RNA PCR kit (part number N808-0017) with M-MLV, the recommended temperature for reverse transcription is 42°C. It is not recommended that reverse transcription be done at a higher temperature with this enzyme because it will start to lose significant activity. If it is necessary to do reverse transcription at a higher temperature due to strong secondary structure or high G+C content, the recommended enzyme to use would be rTth DNA polymerase, which is included in the GeneAmp Thermostable rTth Reverse Transcriptase RNA PCR kit (part number N808-0069).