85-1452-64　［取扱停止］Reverse Transcriptase, M-MLV (M-MLV-RT) Recombinant 2.5KU　R1991-01

特徴

• 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).
• 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, 0.2ug radom hexamer,1-5ug RNA, 200units M-MLV RT. The reaction volume was 20µl and the incubation was 45 min at 42oC.
• 5X Reaaction Buffer:250mM Tris-HCl (pH8.3), 375mM KCl, 15mM MgCl2, and 50mM DTT.
• Applications:1. 1st strand cDNA synthesis
• 2. Primer extension
• 3. RT-PCR.
• Stability:Stable for 5 days at 10oC, for longer period of time store at -20oC. Please avoid freeze-thaw cycles.