High-efficiency stable DNA transfection using cationic detergent and glycerol.
We show that a brief exposure to a simple cationic detergent, in combination with a glycerol 'shock', can result in high frequencies of stable DNA transfection into mammalian cells. Using both Chinese hamster and human cell lines, frequencies of 1 per 1000, or even 1 per 100, viable cells can be achieved readily after optimizing the transfection conditions in a simple buffer. The mechanism of DNA transfer is proposed to differ from that for polycationic liposomes: as well as supplying a hydrophobic counterion, CDAB may promote a conformational change in DNA, allowing the DNA to be readily taken up after the glycerol shock to the cell.
Li
S
S
DNA Repair and Mutagenesis Group, MRC Radiation and Genome Stability Unit, Harwell, Oxfordshire, England.
Thacker
J
J
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D002593:Cetrimonium Compounds; D011232:Chemical Precipitation; D006224:Cricetinae; D003902:Detergents; D005990:Glycerol; D006801:Humans; D013997:Time Factors; D014162:Transfection / Q000379:methods*
D002593:Cetrimonium Compounds; D003902:Detergents; D005990:Glycerol