Add a loop full of E.coli culture to sterile Lauria broth in a test tube using a sterile inoculation loop. Incubate the test tube at 37 degrees Celsius for 24 to 36 hours.
Mix 5 milliliters of E. coli solution to 1 milliliter of detergent in a test tube and place in a hot water bath at 55 to 60 degrees Celsius for 10 minutes. This will free the DNA from the bacterial cells. Add papain enzyme extracts to the test tube and place it in the hot water bath for another 20 minutes. This will remove unwanted proteins attached to the DNA.
Add cold alcohol to the solution to create a layer of about 1 centimeter on top of the bacterial solution. Let the solution sit for two to three minutes. The DNA will precipitate out into the alcohol solution.
Grow a monolayer of the 293 T cells in DMEM medium at 37 degrees Celsius. This should be done 24 hours before the transfection process.
Mix 10 micrograms of DNA to 61 microliters of calcium chloride and 0.5 milliliter of distilled water. Add the solution to 0.5 milliliter of HEPES-buffered saline solution, gently mixing while adding.
Add this mixture to the 293 T cell lines and incubate at 30 degrees Celsius for 16 to 24 hours. The positively charged calcium and the negatively charged phosphate combine with the DNA and enter the eukaryotic cells. This method commonly leads to only short-term expression of the transferred genes.