UNIT 10.17A Stable Transfer of Genes into Mammalian Cells
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Immunology
How to Cite
Kingston, R. E. 2001. Stable Transfer of Genes into Mammalian Cells. Current Protocols in Immunology. 12:VI:10.17A:10.17.1–10.17.6.
- Published Online: 1 MAY 2001
- Published Print: DEC 1994
Analysis of gene function frequently requires the formation of mammalian cell lines that contain the studied gene in a stably integrated form. Approximately one in 104 cells in a transfection will stably integrate DNA, so a dominant selectable marker is used to permit isolation of stable transfectants. Appropriate selection conditions for the parental cell line are first determined, and the gene to be studied is then transfected into that cell line with a gene that expresses a selectable marker. The cells are allowed to grow under selection for approximately 10 doublings before individual colonies are picked and expanded into cell lines. This unit includes conditions for six markers commonly used for selection of mammalian cells. An alternative, specialized procedure for transferring immunoglobulin chains into lymphoid cells is presented in Unit Unavailable .