Magnetic Selection of Transfected Cells
To analyze DNA, RNA, or proteins in their natural environment, isolated genomic DNA or vectors are often transferredcells into cells. The DNA will either become integrated into the genome (stable transfections), or it will remain extrachromosomally until it is diluted by cell division or degraded (transient transfection). Stably transfected cells can be selected with the help of a co-expressed gene, that confers resistance to a cytotoxic drug, which is added to the cell culture. For the selection of transiently transfected cells, this kind of selection is impossible, because the phenotypic manifestation of the drug resistance takes too long. However, by the introduction of cytometric markers instead of drug resistance proteins, and direct physical isolation of transfected cells, instead of killing of the non-transfected cells by drug toxicity, a selection of transient transfectants becomes possible.
KeywordsGreen Fluorescent Protein Untransfected Cell Transfection Procedure Magnetic Cell Sorting Negative Fraction
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- Siebenkotten G, Petry K, Behrens-Jung U, Miltenyi S, and Radbruch A (1997) Employing surface markers for the selection of transfected cells. In: Recktenwald D and Radbruch A (eds.) Cell Separation Methods and Applications. Marcel Dekker Inc. New York, pp 271–281Google Scholar