What is calcium phosphate transfection?

What is calcium phosphate transfection?

Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. A DNA−calcium phosphate co-precipitate forms, which adheres to the cell surface and is taken up by the cell, presumably by endocytosis. Glycerol shock may increase the uptake of DNA in some cell types.

What is the calcium phosphate method?

Abstract. The calcium phosphate transfection is a widely used method for introducing foreign DNA plasmids into cells. Mechanisms underlying this transfection method are not yet defined; however, DNA-calcium phosphate precipitates are internalized by the cells and DNA is efficiently expressed in almost all cell types.

What is calcium phosphate precipitation?

The principle of calcium phosphate co-precipitation involves mixing DNA with calcium chloride in a buffered saline/phosphate solution to generate a calcium-phosphate–DNA co-precipitate, which is then dispersed onto cultured cells.

How does DNA and calcium phosphate co precipitate?

Mix DNA with calcium chloride and add in a controlled manner to a buffered saline/phosphate solution. Incubate at room temperature to generate a precipitate of extremely small, insoluble particles containing condensed DNA. Add the DNA-calcium phosphate co-precipitate to cells, which adhere to the cell membrane.

When was calcium phosphate co-precipitation first used?

Calcium phosphate co-precipitation has been a popular transfection method since its introduction in the early 1970s (Graham and van der Eb, 1973) because the components it requires are easily available and inexpensive.

Can a cell be cytotoxic to calcium phosphate?

However, calcium phosphate co-precipitation is prone to variability due to its sensitivity to slight changes in pH, temperature, and buffer salt concentrations, and can be cytotoxic to many types of cell cultures, especially of primary cells.