Recipe

Digestion Medium

Reagent Quantity (for 1 L) Final concentration
NaCl (MW = 58.44) 23.4 g 400 mm
MgCl2·6H2O (MW = 203.3) 26.4 g 130 mm
MgSO4·7H2O (MW = 246.48) 5.4 g 22 mm
KCl (MW = 74.55) 11.9 g 160 mm
CaCl2 (MW = 147.02) 0.3 g 2 mm
2-(N-morpholino)ethanesulfonic acid (MW = 195.2) 1.95 g 10 mm

  • Adjust the pH to 6.5. Filter through a 0.2-μm filter and store at 4°C. Just before use, add 1% (w/v) cellulose (Sigma) and 3 U/mL alginate lyase (Boyen et al. 1990).

    REFERENCES

    1. ↵
      1. Boyen C,
      2. Kloareg B,
      3. Polne-Fuller M,
      4. Gibor A
      Boyen C, Kloareg B, Polne-Fuller M, Gibor A. 1990. Preparation of alginate lyases from marine molluscs for protoplast isolation in brown algae. Phycologia 29: 173–181.
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    1. doi:10.1101/pdb.rec068726 Cold Spring Harb Protoc 2012: pdb.rec068726- © 2012 Cold Spring Harbor Laboratory Press
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