Product Overview :
Reverse Transcriptase Assay Kit is a convenient, efficient and inexpensive assay for measuring reverse transcriptase activity. The key to this method is our Green dsDNA quantitation reagent, which preferentially detects dsDNA or RNA-DNA heteroduplexes over single-stranded nucleic acids or free nucleotides. In the assay, reverse transcriptase activity in a biological sample generates long RNA-DNA heteroduplexes from a mixture of a long poly(A) template, an oligo-dT primer and dTTP. The RNA-DNA heteroduplexes formed are then detected by the Green reagent. In less than an hour, samples can be read in a fluorometer or microplate reader with filter sets appropriate for fluorescein (FITC).
Storage :
Upon receipt, the kit should be stored at -20°C, protected from light. The 20X TE buffer and the lambda DNA standard are best stored at 4°C; however, either may be frozen for longterm storage. Stored properly, the kit's components should remain stable for at least six months. Allow reagents to warm to room temperature before opening the vials.
Kit Components :
Green dsDNA quantitation reagent (Component A), 0.55 mL of 400X dye in DMSO
20X TE buffer (Component B), 12 mL of 200 mM Tris HCl, 20 mM EDTA, pH 7.5
Lambda DNA standard (Component C), 0.55 mL of 100 µg/mL DNA in TE buffer
Poly(A) ribonucleotide template (Component D), 55 µL of 1 mg/mL template in 100 mM TrisHCl, 0.5 mM EDTA, pH 8.1. The template is approximately 350 bases long
Oligo d(T)16 primer (Component E), 55 µL of 50 µg/mL primer in 100 mM TrisHCl, 0.5 mM EDTA, pH 8.1
Polymerization buffer (Component F), 22.5 mL of 60 mM TrisHCl, 60 mM KCl, 8 mM MgCl2, 13 mM DTT, 100 µM
dTTP, pH 8.1
EDTA (Component G), 2.5 mL of a 200 mM solution in water
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