High efficiency gene transfer by multiple transfection protocol

Histochem J. 1999 Apr;31(4):241-3. doi: 10.1023/a:1003598614323.

Abstract

A high efficiency transfection protocol employing a common polycationic lipid is described. Using LipofectAMINE, a widely used transfection reagent, we transfected 293T cells with a plasmid harboring the beta-galactosidase (beta-gal) gene. The transfection efficiency was determined by direct staining for X-gal. The conventional transfection protocol achieved an efficiency of <40% while our protocol, which employs the repetition of transfection a few times, achieved an efficiency of approximately 80%. Thus, a dramatic increase in transfection efficiency can be obtained by simply repeating transfection with the use of a common polycationic lipid. This method will be useful in many molecular biological experiments.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • COS Cells
  • Cation Exchange Resins*
  • Cells, Cultured
  • Drug Carriers
  • Gene Transfer Techniques
  • Lipids*
  • Transfection / drug effects
  • Transfection / methods*
  • beta-Galactosidase / genetics*

Substances

  • Cation Exchange Resins
  • Drug Carriers
  • Lipids
  • Lipofectamine
  • beta-Galactosidase