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 synthesis of longer cDNAs (>7kb). Source: Recombinant protein corresponding to MMLV, expressed in E. coli strain. Applications: 1. 1st strand cDNA synthesis. 2. Primer extension. 3. RT-PCR. Standard cDNA Synthesis Conditions: 50mM Tris-HCl, pH 8.3, 75mM KCl, 3mM MgCl2, 10mM DTT, 1.0mM each dATP, dGTP, dCTP, and dTTP, 0.2mg radom hexamer,1-5mg RNA, 200units M-MLV RT. The reaction volume was 20ml and the incubation was 45 min at 42oC. 5X Reaction Buffer: 250mM Tris-HCl, pH 8.3, 375mM KCl, 15mM MgCl2, 50mM DTT. Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 1nmol of deoxyribonuc