OPTIMIZATION OF A TRANSIENT TRANSFECTION ASSAY IN COS-1 CELLS

Authors
  1. Nagata, L.P.
Corporate Authors
Defence Research Establishment Suffield, Ralston ALTA (CAN)
Abstract
A transient transfection assay was developed in cos-1 cells to evaluate the level of expression of genes cloned into eucaryotic expression vectors containing the SV40 origin of replication. Transfection yields were quantitated using an indicator plasmid containing the beta-galactosidase gene. Using this plasmid, individual transfected cells could be visualized with a stain containing a chromogenic substrate for beta-galactosidase. In a comparison between two liposome reagents, Lipofectin (TRADEMARK) and TransfectACE (TRADEMARK), the most efficient method of transient transfection for the COS-1 cells was found to be TransfectACE (TRADEMARK). Conditions were optimized for use with this reagent, and transfection efficiencies of 20-25% were attained. The cell permeabilization techniques, scrape-loading and streptolysin-O were also examined alone or in conjunction with TransfectACE (TRADEMARK). Neither of these methods enhanced the efficiency of transfection observed with TransfectACE (TRADEMARK).
Report Number
DRES-M-1409 — Memorandum
Date of publication
01 Apr 1993
Number of Pages
31
DSTKIM No
93-02740
CANDIS No
131627
Format(s):
Hardcopy;Originator's fiche received by DSIS

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