Calcium phosphate transfection method is a very efficient means of introducing DNA into cells in many cell systems. This method works best in cell lines that are 1) highly transformed and 2) adherent. Find here calcium phosphate transfection protocols.
Calcium Phosphate Transfection Method This calcium phosphate transfection method works best in cell lines that are 1) highly transformed and 2) adherent (Hela, U2OS, SAOS2, AdAH, NPC-KT and obtain from 20% to 100% transfection efficiency depending
on the cell line). Works well for transient experiments but precautions should be used in the design and interpretation of experiments based on the discussion below. Also works
very well for generating stable cell lines. This method is quite sensitive to the amount of input plasmid.
Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs Calcium phosphate forms an insoluble precipitate with DNA, which attaches to the cell surface and is taken into the cells by endocytosis. The protocol is easily adapted for use with other types of cells, both adherent and nonadherent. This protocol is a modified version of a method published by Jordan et al. (1996) who rigorously optimized calcium-phosphate-based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells.
Transfecting Cultured Hippocampal Neurons with an Actin-GFP Plasmid This protocol describes two transfection methods for expressing GFP-tagged actin in primary neurons. The lipid reagent DOTAP (Roche Diagnostics) method produces actin-GFP-expressing hippocampal neurons that survive well during long periods in culture. The calcium phosphate method can be used to transfect neurons that have already been growing on coverslips in vitro. Transfected cells suitable for imaging can be obtained in cultures up to 15 days in vitro.
Transfection of Hippocampal Neurons with Plasmid DNA Using Calcium Phosphate Coprecipitation This protocol describes transfection of plasmid DNA into primary hippocampal neurons using DNA/calcium-phosphate (CaPO4) coprecipitation. The precise pH of the transfection medium and the incubation time of cells with the coprecipitate are critical for reproducible and efficient transfection. Once these parameters are optimized for a given plasmid, the method is easily adapted for transfection of other established cell lines.