Unit

UNIT 9.2 Transfection Using DEAE-Dextran

  1. Tod Gulick

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0902s40

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Gulick, T. 2001. Transfection Using DEAE-Dextran. Current Protocols in Molecular Biology. 40:I:9.2:9.2.1–9.2.10.

Author Information

  1. Massachusetts General Hospital, Charlestown, Massachusetts

Publication History

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

Abstract

Transfection of cultured mammalian cells using diethylaminoethyl (DEAE)-dextran/DNA can be an attractive alternative to other transfection methods in many circumstances. The major advantages of the technique are its relative simplicity and speed, limited expense, and remarkably reproducible interexperimental and intraexperimental transfection efficiency. Disadvantages include inhibition of cell growth and induction of heterogeneous morphological changes in cells. Furthermore, the concentration of serum in the culture medium must be transiently reduced during the transfection. In general, DEAE-dextran DNA transfection is ideal for transient transfections with promoter/reporter plasmids in analyses of promoter and enhancer functions, and is suitable for overexpression of recombinant protein in transient transfections or for generation of stable cell lines using vectors designed to exist in the cell as episomes. This unit presents a general description of DEAE-dextran transfection, as well as two more specific protocols for typical experimental applications. The Basic Protocol is suitable for transfection of anchorage-dependent (attached) cells. For cells that grow in suspension, electroporation or lipofection is usually preferred, although DEAE-dextran-mediated transfection can be used.