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 DESCRIPTION  
 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 enzyme requires a primer to intiate synthesis and Mg2+ or Mn2+ for activity.  Compared to AMV Reverse Transcriptase, this enzyme has a much weaker 5' ¡æ 3' ribonuclease H activity, which allows the syntesis of longer cDNAs (>7kb).  
   
 SOURCE  
 Recombinant E. coli strain.  
   
 APPLICATIONS  
 ¡Ü 1st strand cDNA synthesis.  
 ¡Ü Primer extension.  
 ¡Ü RT-PCR.  
   
 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 37¡É, using poly(A)-oligo(dT)12-18 as the template-primer.  
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 STANDARD UNIT ASSAY CONDITIONS  
 50mM Tris-HCl (pH8.3), 75mM KCl, 3mM MgCl2, 10mM DTT, 0.5mM each dATP, dGTP, dCTP, and dTTP, 1-10 ¥ìCi of [¥á32P]dCTP added as a tracer, 10¥ìg/ml oligo (dT)12-18, 20¥ìg/ml mRNA, 200units M-MLV RT.  The reaction volume was 20¥ìl and the incubation was 60 min at 37¡É.  
   
   
 CONCENTRATION: 200 units/¥ìl.  
   
   
 5X REACTION BUFFER  250mM Tris-HCl (pH8.3), 375mM KCl, 15mM MgCl2, and  50mM DTT.  
   
   
 STORAGE CONDITIONS  
 0.1M KPO4 (pH7.2), 0.2% Triton X-100, 2mM DTT, and 50% glycerol.  Store at -20¡É.¡¡   
   
   
 REFERENCES  
 1. Sambrook, J., et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.  
 2. Gerald, G. F. and D'Alessio, J. M. (1993) In Methods in Molecular Biology, Vol. 16, Humana Press, NJ.   |