Quality level: 200
Qualification: for molecular biology
Sterility: sterile; 0.2 μm filtered
Form: solution
Use
- kit enough for 160 transfections (6 cm dishes)
- kit enough for 400 transfections (3.5 cm dishes)
- kit enough for 80 transfections (10 cm dishes)
Application(s): transfection: adequate
Sent in: dry ice
Storage temperature: −20 ° C
General description
Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. This technique has been used to obtain transient and stable transfections in a wide variety of cell types.
Request
The calcium phosphate transfection kit has been used:
to allow transfection
to transfect Hek293T cells
to transfect H29D cells
Suitable for transient and stable transfection of DNA in cultured mammalian cells. The following cells have been successfully transfected using the calcium phosphate method:
- BASS
- Intestinal melanoma cells
- CHO K1
- COS-7
- Fibroblasts (human embryonic, neo derm)
- HEK293
- Eh 7
- IMR-90
- LLC (Lewis lung carcinoma)
- NIH3T3
- PC-12
- PCI-13
- SH-Sy5Y
- SK-Hep-1
- T47D
Features and Benefits
- Suitable for transient and stable transfection
- Reproducible for a wide range of cell types
- Widely referenced
- Cheap
Components
The Calcium Phosphate Transfection Kit contains:
5 ml of 2.5 M CaCl2 (C2052)
25 ml 2x HEPES buffered saline (H1012)
25ml molecular biology grade water (W4502)
Beginning
The procedure is based on slowly mixing HEPES-buffered saline containing sodium phosphate with a CaCl2 solution containing DNA. A DNA-calcium phosphate co-precipitate forms adhere to the cell surface and are taken up by the cell, presumably by endocytosis.
Other notes: The protocol was developed for transient transfection of CHO cells using the plasmid pSV40-CAT as a reporter gene.