Moloney Mouse Leukemia Virus Reverse Trancscriptase (Recombinant)

Recombinant Moloney Murine Leukemia Virus Reverse Trancscriptase

Source: Recombinant E. coli strain

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 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).

Applications: 1. 1st strand cDNA synthesis. 2. Primer extension. 3. RT-PCR.

Physical Appearance: Sterile Filtered clear solution (200U/ l).

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, 0.2 g radom hexamer,1-5 g RNA, 200units M-MLV RT. The reaction volume was 20 l and the incubation was 45 min at 42oC.

Concentration: 200 units/ l.

5X Reaaction Buffer: 250mM Tris-HCl (pH8.3), 375mM KCl, 15mM MgCl2, and 50mM DTT.

Formulation: 50mM Tris-HCl, 0.1M NaCl, 0.1% Triton X-100, 2mM DTT, 0.1mM EDTA and 50% glycerol.

Stability: Stable for 5 days at 10oC, for longer period of time store at -20oC. Please avoid freeze-thaw cycles.

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.

Usage: BioWORLD\\\'s products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.