Unit

UNIT 4.6 Selection of Transfected Mammalian Cells

  1. Richard Mortensen1,
  2. Jonathan D. Chestnut2,
  3. James P. Hoeffler (magnetic bead selection)2,
  4. Robert E. Kingston3

Published Online: 1 MAY 2001

DOI: 10.1002/0471142301.ns0406s00

Current Protocols in Neuroscience

Current Protocols in Neuroscience

How to Cite

Mortensen, R., Chestnut, J. D., Hoeffler, J. P. and Kingston, R. E. 2001. Selection of Transfected Mammalian Cells. Current Protocols in Neuroscience. 00:4.6:4.6.1–4.6.20.

Author Information

  1. 1

    Brigham and Women's Hospital, Boston, Massachusetts

  2. 2

    Invitrogen Corporation, Carlsbad, California

  3. 3

    Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: SEP 1997

Abstract

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 (the efficiency can vary depending on the cell type). Therefore, a dominant, selectable marker is used to permit isolation of stable transfectants. In the first part of this unit, the procedure for determining selection conditions and the resulting stable transfection is presented and the most commonly used selectable markers are discussed. The second protocol includes conditions for thirteen markers commonly used for selection of mammalian cells. A third protocols describes selection of transfected cells from the total population soon after transfection with plasmids that express both the gene of interest and a selection tag. Optimization of transfection conditions can be facilitated by a simple staining assay detailed in a support protocol.