ELISA REDscript(TM) M-MLV Reverse Transcriptase (H-)

REDScript(TM) M-mLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (MmLV RT) is an RNA-dependent DNA polymerase that syntheQuantitys 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/μl Component: M-mLV （200U/μl） 5×Buffer （with DTT） Package： BµLk Features: Weak RNaseH activity High cDNA yield Storeage: -20℃ Unit Definition: One unit of REDscript(TM) M-mLV RT catalyzes the 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 synthesis 1) 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) 1μl Total RNA (10ng-5μg) or mRNA(1-500ng) xμl dNTP (10mM each) 1μl DEPC ddH2O (14-x)μl 2) Gently mix and incubate 10 Min at 70℃ then chill on ice for 2-10min. 3) Centrifµge for a few seconds then Put the tube into ice and add the next composition : 5×RT Buffer 4μl RNasin (40U/μl) 1μl 5) 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) Centrifµge 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.