P buffer

P (protoplast) buffer (Okanishi et al., 1974; Hopwood and Wright, 1978)

Make up the following basal solution:

• 103 g Sucrose
• 0.25 g K₂SO₄
• 2.02 g MgCl₂.6H₂O
• 2 ml Trace element solution (Same as R2 and R2YE)
• 800 ml dH₂O

Dispense in 80 ml aliquots and autoclave. Before use, add to each flask in order:

• 1 ml KH₂PO₄ (0.5%)
• 10 ml CaCl₂.2H₂O (3.68%)
• 10 ml TES buffer (5.73%, adjusted to pH7.2)