Unit

UNIT 12.3 Preparation of Adenovirus-Polylysine-DNA Complexes

  1. Matt Cotten1,
  2. Adam Baker1,
  3. Max L. Birnstiel1,
  4. Kurt Zatloukal2,
  5. Ernst Wagner3

Published Online: 1 MAY 2001

DOI: 10.1002/0471142905.hg1203s11

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Cotten, M., Baker, A., Birnstiel, M. L., Zatloukal, K. and Wagner, E. 2001. Preparation of Adenovirus-Polylysine-DNA Complexes. Current Protocols in Human Genetics. 11:12.3:12.3.1–12.3.33.

Author Information

  1. 1

    Institute for Molecular Pathology, Vienna, Austria

  2. 2

    University of Graz Medical School, Graz, Austria

  3. 3

    Boehringer Ingelheim R & D Vienna, Vienna, Austria

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: NOV 1996

Abstract

This unit describes preparation of adenovirus-polylysine-DNA complexes, which is useful for transfection of DNA into a variety of cell types. A DNA complex is prepared with biotinylated adenovirus and streptavidin-polylysine, coupled to transferrin, and used to transfect cells. Several support protocols describe methods for adenovirus growth and purification, biotinylation, inactivation with psoralen, and quantitation of the adenovirus particles. Additional support protocols descibes preparation of streptavidin-polylysine and transferrin-polylysine, necessary for the basic procedure. The DNA used for transfection must be free of lipopolysaccharide (LPS), and two methods for removing LPS are described. A more direct polylysine-virus linkage that is simple and requires no exotic reagents can be used for transfection. This protocol requires polylysine-modified adenovirus, prepared as described. An alternate protocol describes transfecting cells with free virus and DNA condensed with a polycation.