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.
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.
Storage and Stability:May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
仕様
Size：2500U
Source Antigen：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, 50% glycerol.
