L buffer

L (lysis) buffer (Thompson et al. 1982)

Mix the following sterile solutions:

• 100 ml Sucrose (10.3%)
• 10 ml TES buffer (5.73%, adjusted to pH7.2)
• 1 ml K₂SO₄ (2.5%)
• 0.2 ml Trace element solution
• 1 ml KH₂PO₄ (0.5%)
• 0.1 ml MgCl₂ . 6H₂O (2.5M)
• 1 ml CaCl₂ (0.25M)

This stock solution keeps indefinitely. Just before use dissolve lysozyme in a sample of the solution at a concentration of 1 mg/ml and sterilise by filtration.