QuantiFluor® dsDNA System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Exposure to light will reduce the sensitivity of the assay. Store QuantiFluor® dsDNA Dye and working solution protected from light

Upstream tips
Exposure to light will reduce the sensitivity of the assay. Store QuantiFluor® dsDNA Dye and working solution protected from light

Publication protocol

Cell proliferation assays
Cell proliferation on 3D silk scaffolds was measured by total DNA quantification using a QuantiFluor dsDNA system (Promega, Madison, WI) according to the manufacturer's protocol. Briefly, 5 × 104 unmodified NIH3T3 cells, EMT6 cells, or a mix of both cell lines at a 9:1 ratio was seeded onto the scaffolds. At days 1, 5, 10 or 14, the cells on the scaffolds were washed with PBS and lysed in 750 μL Cell Lytic M reagent (Sigma, St. Louis, MO) for 1 h with shaking. Lysates were frozen at -20°C. For the assay, lysates were diluted 10 × with Cell Lytic Reagent M and mixed at a 1:1 ratio with the supplied working solution of dsDNA dye. After a 5 min incubation at RT, fluorescence was measured using a Victor X3 Multimode Plate Reader (PerkinElmer, Waltham, MA) controlled by the PerkinElmer 2030 Workstation software (PerkinElmer, Waltham, MA). The excitation wavelength of 504 nm and emission of 531 nm were used. The experiment was repeated three times in triplicate.

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Manufacturer protocol

Download the product protocol from Promega for QuantiFluor® dsDNA System below.

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