Abstract
FACS-Gal is a system that enables the fluorescence activated cell sorter (FACS) to sensitively assay expression of the E. Coli lacZ (β-galactosidase, β-gal) reporter gene, and sort viable cells based on the levels of expression of this enzyme.1,2 The system depends on the enzymatic conversion of the nonfluorescent β-gal substrate fluorescein di-β-galactopyranoside (FDG), into the fluorescent molecule fluorescein. FACS-Gal effectively combines a selectable marker with a reporter gene and in the combination produces novel experimental possibilities. The system is an approach to applying the analytical and sorting capabilities of the FACS to solving problems in molecular biology. FACS-Gal and its variants have been used with cells from a variety of organisms including E. Coli,3 yeast,3 Drosophila,4 transgenic mice,5–7 and most frequently with mammalian cell lines.8–13 For a more extensive list of references using the FACS-Gal assay see.14 This review focuses on the technical basics of FACS-Gal, how to most productively use the system, and the strengths and weaknesses of this system in comparison to other presently available alternatives.
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References
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Fiering, S. (2000). FACS-Gal: Flow Cytometric Assay of β-galactosidase in Viable Cells. In: Diamond, R.A., Demaggio, S. (eds) In Living Color. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-57049-0_20
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DOI: https://doi.org/10.1007/978-3-642-57049-0_20
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