DESCRIPTION
Moloney Murine Leukemia Virus (M-MuLV) reverse transcriptase is a RNA-dependent DNA polymerase. This enzyme can synthesize a complementary DNA strand initiating from a primer using either single-stranded RNA or DNA template. The enzyme lacks RNAseH activity.

SOURCE
E.coli strain that carries a plasmid with the cloned and modified M-MuLVRT gene with deleted RNAseH coding part.

CONCENTRATION
100 units/µl

UNIT DEFINITION
One unit is the amount of enzyme required to incorporate 1 nmol of dTTP into an acid insoluble form in 10 minutes at 37°C using poly(rA)-oligo(dt) 10-20 as template primer.

STORAGE BUFFER
50 mM Tris-HCl pH 8.3, 1 mM EDTA, 0.1 mM DTT, 0.1 mM NaCI, 0.1% Triton X-100, 50% glycerol

5X REACTION BUFFER
250 mM Tris-HCI pH 8.3, 15 mM MgCl₂, 400 mM KCl

Add to buffer: dNTPs (end concentration 2 mM), MnCl₂ (end concentration 2-4 mM) and DTT (end concen-tration 10 mM). Incubate at 37°C.

EXTRA SOLUTION
25 mM MnCl2 100 mM DTT

UNIT ASSAY CONDITION
20 mM Tris-HCI pH 8.0, 2 mM MnCl2 , 100 mM KCl, 1 mM DTT, 0.6 mM poly rA, 0.1 mM poly(dT)10-20; 0.5 mM dTTP (3H) - 0.5-5 units of enzyme

QUALITY ASSURANCE
GeneScript™ reverse transcriptase is tested for its ability to synthesize full length cDNA from 4kb RNA.