Invitrogen™

M-MLV Reverse Transcriptase (200 U/µL)

Catalog number:  28025021

Related applications:

Reverse Transcription

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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. Compared to AMV RT, Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) lacks DNA endonuclease activity and has a lower RNase H activity. Features of this enzyme:

Thermostability—optimal activity at 37°C
Size of cDNA—M-MLV can be used to synthesize first-strand cDNA up to 7 kb
Applications—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 definition
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.

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

Concentration: 200 U⁄µl
Format: Tube(s)
Includes: 1 × 1mL M-MLV RT (200U/μL); 1mL 5X First Strand Buffer [250mM Tris-HCl (pH 8.3), 375mM KCl, 15mM Magnesium Chloride]; 500μL 100mM DTT; 200,000 units
No. of Reactions: 1000 reactions
Quantity: 1000 reactions
Sensitivity: Low
Enzyme Function: RNA dependent DNA Polymerase
Size (Final Product): 7 kb or less
Optimal Reaction Temperature: 37° C
Reverse Transcriptase: M-MLV
Ribonuclease H Activity: Reduced
Shipping Condition: Wet Ice
Template: ssDNA, ssRNA, RNA-DNA Hybrid

Contents & storage

Contains:
• 1 × 1 mL M-MLV RT (200 U/µl)
• 1 ml 5X First Strand Buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM Magnesium Chloride]
• 500 µl 100 mM DTT

Store at -20°C. Guaranteed stable for 6 months when properly stored.