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

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

Catalog No. 28025013
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28025013 200 Reactions
28025021 1,000 Reactions
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Catalog No. 28025013 Supplier Invitrogen™ Supplier No. 28025013
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Description

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.

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

Order Info

Shipping Condition: Wet ice

Specifications

Concentration 200 U/μL
Content And Storage

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

Store at –20°C.
Format Stand-alone Enzyme
Reaction Speed 50 min.
Technique Reverse Transcription
Optimal Reaction Temperature 37°C
Quantity 40,000 units
Reverse Transcriptase M-MLV
Ribonuclease H Activity Yes
Shipping Condition Wet Ice
Final Product Type First-Strand cDNA
No. of Reactions 200 Reactions
Reaction Format Separate components
Reagent Type Reverse Transcription
Size (Final Product) Up to 7 kb
Starting Material RNA
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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.

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

The optimal temperature for for MMLV is 42 degrees C. The optimal temperature for SuperScript II RT is 42 degrees C, and can be used up to 50 degrees C. 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 IV RT is 50 degrees C, but can be used up to 65 degrees C.

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

Moloney Murine Leukemia Virus (M-MLV) Reverse Transcriptase is not well-suited for elevated temperatures. Typically, M-MLV RT operates optimally at temperatures around 37 degrees C to 42 degrees C.

To perform reverse transcription at elevated temperatures (e.g., 50 degrees C or higher), consider using thermostable reverse transcriptases such as SuperScript IV or Maxima H Minus, which are engineered to perform efficiently at higher temperatures.

For Research Use Only. Not for use in diagnostic procedures.