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).\n\nUnit Definition: \nOne 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.\n\nStandard cDNA Synthesis Conditions: \n50mM 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 20microl and the incubation was 45 min at 42oC.\n\n5X Reaaction Buffer: \n250mM Tris-HCl (pH8.3), 375mM KCl, 15mM MgCl2, and 50mM DTT.\n\nApplications:\n1. 1st strand cDNA synthesis.\n2. Primer extension.\n3. RT-PCR.\n\nStability: \nStable for 5 days at 10oC, for longer period of time store at -20oC. Please avoid freeze-thaw cycles.

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SPECIFICATIONS

Catalog Number

R1991-01

Size

2.5KU

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.

References

1. Sambrook, J., et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.\n2. Gerald, G. F. and D'Alessio, J. M. (1993) In Methods in Molecular Biology, Vol. 16, Humana Press, NJ.\n\n1. Study of MMLV RT- binding with DNA using surface plasmon resonance biosensor.\n\nActa Biochim Biophys Sin (Shanghai) 2005 Sep;37(9):634-42\n\n2. Incorporation of (alpha-P-borano)-2',3'-dideoxycytidine 5'-triphosphate into DNA by drug-resistant MMLV reverse transcriptase and Taq DNA polymerase.\n\nNucleosides Nucleotides Nucleic Acids 2003 May-Aug;22(5-8):1651-5\n\n3. Sensitivity of tyrosinase mRNA detection by RT-PCR: rTth DNA polymerase vs. MMLV-RT and AmpliTaq polymerase.\n\nBiotechniques 1996 Apr;20(4):592-600