What is the procedure for establishing a new protocol for the electroporation of eukaryotic cells?
1. Check whether the cells will tolerate the hypoosmolar buffer. After 30 minutes’ incubation, 90 % of the cells should survive, otherwise mixtures of hypoosmolar and isoosmolar buffer will have to be prepared until the desired survival rate is achieved. 2. Cell size can be used as a guide to the voltage to be used. A good approach is to make a rising series of voltages in 50 V increments. The time constant is usually 40 – 100 µs. Detailled information are included in the “General Protocol” and “Optimization Protocol” or in the Basic Application Manual for Electroporation which can be found here.
Related Questions
- Is it necessary following the electroporation to incubate eukaryotic cells on ice for 10 minutes, as is frequently recommended in the protocols?
- What is the procedure for establishing a new protocol for the electroporation of eukaryotic cells?
- What is the maximum number of eukaryotic cells which can be used for an electroporation?
