Overview of our system

Vector information

Protocols

Protoplast isolation

Gene transfection

Luciferase assay

Western blotting

Results

Transfect efficiency

Gene expression levels of dual expression vectors

Protein interaction

Acknowledgements

<Back to Home>

Updated 96-well plate protoplasts transfection (11/30/09)

We developed a 96-well plate assay method, with Arabidopsis protoplasts, by adapting the method Sheen and colleagues reported (He, P. et al, J, 2007). We found that vortexing a 96-well plate after adding a polyethylene glycol (PEG) solution and centrifuging the plate after each washing step (see materials and methods for the details) allowed higher-efficient transformation with less number of protoplasts, compared to the original method, in our hands

Instrument

8-channel digital pipette	Centrifuge machine	U96 MicroWell Plates ( Nalge Nunc International)
digital vortex	Thermomixer R (Eppendorf AG,)	Washer ELx405TM Microplate Washer (BioTek)

Material

polyethlyen glycol (PEG) solution

40%(w/v) PEG4000 (Fluka, Buchs SG, Switzerland)
0.2 M mannitol
0.1 M CaCl₂

Protocol

Download from here

Results

Transfect efficiency and gene expression