HeLa Nuclear Extract

Publisher Summary This chapter focuses on HeLa nuclear extracts. The chapter presents a method that is used for the growth of HeLa cells. HeLa S3 cells (human epithelioid carcinoma, cervix) were obtained from ATCC stocks. The cells are adapted to grow in Joklik's MEM medium; containing 5% defined Bovine Calf Serum, supplemented with penicillin-streptomycin and NaHCO3 at 37°C. The cells were grown in suspension and stirred using magnetic bars at medium speed. All flasks should be tightly sealed in order to keep the carbon dioxide (CO2 ) at appropriate levels. HeLa cells are grown at an ideal density of 5–8 × 105/ml (minimum density should be 3 × 105/ml in 20 l of medium. HeLa cells double approximately every 24 h, and to keep a density of 5–8 × 105/ml, half of the volume of the cell suspension should be discarded daily and the same amount of fresh medium should be added until expansion is required. This chapter also discuses the extract preparation including: materials, methods, and nuclear extraction.