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Table 1 Troubleshooting

From: In vivo monitoring of plant small GTPase activation using a Förster resonance energy transfer biosensor

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