Skip to main content

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