Problem | Possible reason | Solution |
---|---|---|
Low protoplast yield and low transfection efficiency | Condition of suspension cells | Suspension cell condition is the most important factor for obtaining successful results. If you have problems with a transfection, prepare new suspension cells from your stock or rice seeds |
Insufficient cellulase digestion | Optimize the period of cellulase treatment or prepare fresh cellulase solution | |
Problem with transfection reagents | Prepare new transfection reagents. PEG solution can not be stored, and you should prepare fresh PEG for each experiment | |
Low DNA quality | Use an animal transfection grade of DNA purification kit | |
Decay of FRET signal during imaging | Photobleaching by the excitation | Minimize exposure time and duration of laser excitation |