REDScript(TM) M-MLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (MMLV RT) is an RNA-dependent DNA polymerase that synthesizes the complementary DNA (cDNA) first strand from a single-stranded RNA template to which a primer has been hybridized. M-MLV RT will also extend primers hybridized to single-stranded DNA. Second strand cDNA synthesis can be achieved from some mRNA templates without an additional DNA polymerase. The difference between this to the general M-MLV RT is that the capacity to endure the heat is enhanced. It can remain the 100% activity at 50℃, it can also keep more than 75% activity at even at 55℃. Upper panel: Template: Mouse total RNA, Reverse transcription at 50℃ Lane 1-2: REDscript(TM) M-MLV 200U Lane 3-4 :invitrogen SuperScript® III Lane 5-6:Takara m-mlv Lane M: Marker Lower panel: Template: Mouse total RNA, Reverse transcription at 55℃ Lane 1-2:Takara m-mlv Lane 3-4: REDscript(TM) M-MLV Lane 5-6:200U invitrogen SuperScript® III Lane M: Marker Source: Recombination of E.coli containing Moloney murine leukemia virus reverse transcriptase gene from clone of Moloney murine.Concentration: 200U/μlComponent: M-MLV （200U/μl） 5×Buffer （with DTT） Package： BulkFeatures: Weak RNaseH activity High cDNA yieldStoreage: -20℃ Unit Definition: One unit of REDscript(TM) M-MLV RT catalyzesthe incorporation of 1 nmol of dTTP into acidinsoluble material in 10 minutes at 37℃ using oligo(dT)12-18-primed poly(A)n as a template. Applications: The first-strand cDNA synthesis; RT-PCR. Storage Buffer: 20 mM Tris-HCl (pH7.5), 200 mM NaCl，0.25 mM EDTA, 0.01% NP-40(v/v)，2.5 mM DTT,50% glycerol (v/v).5X Reaction Buffer:[5×RT Buffer] 250mM Tris-HCl (pH 8.3), 15mM MgCl2，375 mM KCl，50mM DTT.Recommended Reaction Conditions:The first-strand cDNA synthesis1) Add the following reagents to a RNase free PCR tube at room temperature add the MMLV RT last.Oligo dT12-18 (1μg/μl) or random primer (50-250ng) 1μlTotal RNA (10ng-5μg) or mRNA(1-500ng) xμldNTP (10mM each) 1μlDEPC ddH2O (14-x)μl2) Gently mix and incubate 10 Min at 70℃ then chill on ice for 2-10min.3) Centrifuge for a few seconds then Put the tube into ice and add the next composition :5×RT Buffer 4μlRNasin (40U/μl) 1μl5) Gently mix and incubate at 50℃ for 2 Min（Oligo dT12-18 or sequence especially primer）or at 25℃ for 10 min for the random primer.6) Centrifuge for a few seconds. Add 1μl REDscript(TM) M-MLV RT（200U/μl）Incubate at 50℃ for 50min.7) Inactivate at 70℃ for 10min then get the cDNA.
