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MES(cas 4432-31-9) is used for the isolation, fusion and culture of plant protoplasts

Reagents:

(1) Enzyme solution: 1.25% cellulase, 0.3% pectinase, 0.04% mannitol, 20 mmol /L KCl and 20 mmol /L MES(cas 4432-31-9) were added successively, and the mixture was cooled to room temperature, plus 10mmol /LCaCL2, 5mmol /L β-mercaptoethanol and 0.1% BSA, 0.45um microporous membrane filtration, the solution was transparent orange. 

(2) PEG solution: 4GpPEG4000 /3 mL deionized water, 2.5 mL 0.8 mol /L mannitol and 1 mL 1 mol /L CaCl2.

Plant protoplasts are "bare cells" surrounded by protoplasts when the cell wall is removed, making it ideal for basic research. The principle of cell wall is mainly composed of cellulose, hemicellulose and pectin, so the use of cellulase, hemicellulase and pectinase can degrade cell wall components, remove the cell wall.

 

 

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