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MES(cas 4432-31-9) used for the preparation of cell protoplasts

1. Materials: mung bean, tobacco seedling leaves, rape or spinach or tobacco. 

2. Reagents:

(W / V) pectinase, 0.7 mol /L mannitol; 10 mmol /L CaCl, 2.2 H2O, 0.7 mmol /L KH2PO4, pH 6.8 to 7.0. 
(2) 13% CPW Washing liquid (mung bean): 27.2 mg / L KH2PO4, 101.0 mg /L KNO3, 1480.0 mg /LCaCl, 2.2 H2O, 246.0 mg /L MgSO4, 0.16 mg /L KI, 0.025 mg /5H2O, 13% (w /v) mannitol, pH 6.0.
(3) 0.1% MES(cas 4432-31-9) or trimethylolaminomethane solution: 20% sucrose, pH 6.0. 
(4) 0.02% diethylfluorescein (FDA): 5mg FDA dissolved in 1ml acetone, stored at 4 ℃ in the dark, use 0.22ml FDA storage solution by adding 5ml0.65mol /L mannitol. The concentration was 0.01%.
(5) 20% sucrose solution
(6) Sterile distilled water.

 

 

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