| <b>Materials Required but not Supplied</b> RNase Inhibitor, Murine (NEB #M0314) Nuclease-free Water (NEB #B1500) <b>Unit Definition</b> One unit is defined as the amount of enzyme that will incorporate 1 nmol of dTTP into acid-insoluble material in a total reaction volume of 50 μl in 10 minutes at 37°C using poly(rA) - oligo(dT) as template primer with 50 mM Tris-HCI (pH 8.3), 6 mM MgCl 2 , 10 mM dithiothreitol, 0.5 mM [ 3 H]-dTTP and 0.4 mM poly(rA) - oligo(dT)12-18. <b>Reaction Conditions</b> 1X M-MuLV Reverse Transcriptase Reaction Buffer Incubate at 42°C 1X M-MuLV Reverse Transcriptase Reaction Buffer 50 mM Tris-HCl 75 mM KCl 3 mM MgCl2 10 mM DTT (pH 8.3 @ 25°C) <b>Storage Buffer</b> 50 mM Tris-HCl 150 mM NaCl 1 mM DTT 0.1 mM EDTA 50% Glycerol 0.1% NP-40 pH 7.6 @ 25°C <b>Heat Inactivation</b> 65°C for 20 minutes <b>Molecular Weight</b> Theoretical | 78000 daltons |
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