X-Gal

5-bromo-4-chloro-3-indolyl-beta-galactopyranoside

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X-gal is the most widely used chromogenic substrate for β-galactosidase. Enzyme hydrolysis of the substrate yields a dark blue colored precipitate (λmax ~615 nm).

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    • How can I measure the ratio of successful transfections?

      Transfection rates can directly be measured by (co-)transfecting a GFP-expressing plasmid. The percentage of cells that have taken up and expressed the foreign nucleic acid can be visualized with a fluorescence microscope (by green autofluorescence).
      Another way of quantification is to transfect luciferase or beta-galactosidase expressing reporter vectors and to determine the activity of these enzymes as a measure of successful transfection. Several commercial cell based reporter assays are available for this purpose (Firefly and Renilla Luciferase Reporter Assay Kits; GFP reporter Assay Kit; beta-Galactosidase Activity Assay Kit; all available from PromoKine).

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