An Automated Protoplast Transformation System
Efficient plant protoplast production from cell suspension cultures, leaf, and stem tissue allows for single-cell plant biology. Since protoplasts do not have cell walls, they can be readily transformed to enable rapid assessment of regulatory elements, synthetic constructs, gene expression, and more recently genome-editing tools and approaches. Historically, enzymatic cell wall digestion has been both expensive and laborious. Protoplast production, transformation, and analysis of fluorescence have recently been automated using an integrated robotic system. Here we describe its use for bulk protoplast isolation, counting, transformation, and analysis at very low cost for high-throughput experiments.
Key wordsTobacco Protoplasts Transformation Enzymatic digestion High-throughput screening Automation Robotics
This research was supported by Advanced Research Projects Agency-Energy (ARPA-E) Award No. DE-AR0000313, with additional funding and assistance from ThermoFisher.
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