Reverse Transcriptase, M-MLV (M-MLV-RT) Recombinant
MMLV (Molony Murine Leukemia Virus) Reverse Transcriptase is a DNA polymerase that synthesizes a complementary DNA strands from single-stranded RNA, DNA, or an RNA-DNA hybrid as a template. This recombinant enzyme was purified from E.coli. which carried a modified MMLV-RT gene. Compared to AMV Reverse Transcriptase, this enzyme has a much weaker 5'-3' ribonuclease H activity, which allows the syntesis of longer cDNAs (>7kb).
Unit Definition
One unit is defined as the amount of enzyme required to catalyze the incorporation of 1nmol of deoxyribonucleotide into acid-insoluble forms in 10 minutes at 37oC, using poly(A)-oligo(dT)12-18 as the template-primer.
Standard cDNA Synthesis Conditions
50mM Tris-HCl (pH8.3), 75mM KCl, 3mM MgCl2, 10mM DTT, 1.0mM each dATP, dGTP, dCTP, and dTTP, 0.2ug radom hexamer,1-5ug RNA, 200units M-MLV RT. The reaction volume was 20µl and the incubation was 45 min at 42oC.
5X Reaaction Buffer
250mM Tris-HCl (pH8.3), 375mM KCl, 15mM MgCl2, and 50mM DTT.
Applications
1. 1st strand cDNA synthesis. 2. Primer extension. 3. RT-PCR.
Stability
Stable for 5 days at 10oC, for longer period of time store at -20oC. Please avoid freeze-thaw cycles.
Source
Recombinant, E. coli
Form
Supplied as a liquid in 50mM Tris-HCl, 0.1M sodium chloride, 0.1% Triton X-100, 2mM DTT, 0.1mM EDTA and 50% glycerol.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
