Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. This technique has been used to obtain both transient and stable transfections in a wide variety of cell types.
Calcium Phosphate Transfection Kit has been used:
- to enable transfection
- to transfect Hek293T cells
- to transfect H29D cells
Suitable for transient and stable transfection of DNA into cultured mammalian cells. The following cells have successfully been transfected using the calcium phosphate method:
Bowels melanoma cells
Fibroblasts (human embryonic, neo derm)
LLC (Lewis Lung Carcinoma)
Caractéristiques et avantages
- Suitable for transient and stable transfection
- Reproducible for a wide range of cell types
- Widely referenced
The Calcium Phosphate Transfection Kit contains:
5 ml 2.5M CaCl2 (C2052)
25 ml 2x HEPES Buffered Saline (H1012)
25 ml molecular biology grade water (W4502)
The procedure is based on slow mixing of HEPES-buffered saline containing sodium phosphate with a CaCl2 solution containing the DNA. A DNA-calcium phosphate co-precipitate forms, which adheres to the cell surface and is taken up by the cell, presumably by endocytosis.
The protocol was developed for transient transfection of CHO cells using pSV40-CAT plasmid as a reporter gene.