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Published on: December 25, 2015
High yields of DNA-transfer into mouse L-cells by electropermeabilization
Zeitschrift Fur Naturforschung. Section C, Biosciences
|November 1, 1985
Summary
Efficient DNA transfection in mouse cells was achieved using electropermeabilization. This method yields high numbers of neomycin-resistant clones, even with low DNA concentrations, optimizing gene delivery.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- DNA transfection is crucial for genetic modification of cells.
- Electroporation is a common method for introducing DNA into cells.
- Optimizing transfection protocols is essential for efficient gene delivery and stable cell line development.
Purpose of the Study:
- To optimize DNA transfection in mouse L-cells using electropermeabilization.
- To achieve high yields of stably transfected clones with neomycin resistance.
- To investigate the impact of various parameters on transfection efficiency.
Main Methods:
- Electroporation (electropermeabilization) technique was employed for DNA transfection.
- Plasmid pSV 2-neo was used to confer neomycin resistance in transfected cells.
- Optimization involved varying temperature, field parameters, and medium composition.
Main Results:
- High yields of stably transfected L-cells (clones) were obtained.
- Efficient transfection was achieved at relatively low DNA concentrations.
- The study identified optimal conditions for pulse application and resealing.
Conclusions:
- Electroporation is an effective method for high-yield DNA transfection in mouse L-cells.
- Optimized conditions enhance the efficiency of generating neomycin-resistant cell lines.
- Parameters like temperature and medium composition significantly influence transfection success.
