Dilute FluoroVue™ Nucleic Acid Gel Stain 10,000 folds into the TE, TAE, or TBE buffer and mix well. Performing agarose gel electrophoresis following your standard protocol.Ģ. The sensitivity of this method is lower than the in-gel staining method.ġ. Post-staining method is recommended for polyacrylamide electrophoresis, due to the longer time required for running PAGE. Staining after electrophoresis (post-staining) Video cameras and CCD cameras have a different spectral response compared to the black-and-white print film and therefore may not exhibit the same sensitivity. Accumulation of fluorescent dyes on the surface will create a high fluorescent background. Visualize or photograph the gel with UV or blue-light illumination (blue-light is recommended).Ĭlean the surface of the illuminator before and after each use with deionized water. High voltage causes excess heat and affects the dye adversely.ĭuring electrophoresis, the staining dye runs toward the anode, therefore DNA bands with smaller molecular weights may be weaker in intensity due to less staining dye.ĥ. Avoid using high voltage during electrophoresis. The recommended voltage is 4–10 V/cm (distance between anode and cathode). Perform agarose gel electrophoresis (avoid light). After three days the sensitivity will decrease daily.Ĥ. Cool the molten agarose solution until it can be handled by hand and then pour it into gel tray.Ĭasted gels are stable at 4☌ for 3 days in dark. Dilute FluoroVue™ Nucleic Acid Gel Stain 10,000X with the molten gel solution and mix well prior to being poured into the gel.įor example: 4 μl in 40 ml molten agarose solutionģ. Prepare TAE or TBE buffered molten agarose solution.Ĥ0 ml molten agarose solution can cast two mini-gels (5.4 x 5.9 cm) or one landscape-gel (10.9 x 5.9 cm).Ģ. Increased cloning efficiency (blue light)ġ. Sensitivity: 0.14 ng (DNA) or 1 ng (total RNA)Ĭompatibility: suitable to blue or UV light Therefore, it can replace EtBr without the need of changing existing lab imaging systems. When bound to nucleic acids, FluoroVue™ Nucleic Acid Gel Stain has a fluorescent excitation maximum of 250 and 482 nm, and an emission maximum of 509 nm. FluoroVue™ Nucleic Acid Gel Stain offers high sensitivity that is several times greater than EtBr.įluoroVue™ Nucleic Acid Gel Stain is compatible with both conventional UV gel-illumination systems as well as harmless long wavelength blue light illumination systems, like B-BOX™. It is a fluorescent stain which offers highly sensitive detection of double-stranded or single-stranded DNA and RNA in a convenient manner. FluoroVue™ Nucleic Acid Gel Stain (10,000X) is specially designed for in-gel use and is a safer replacement for conventional Ethidium bromide (EtBr), which poses a significant health and safety hazard to its users.
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