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Holtfreter's Solution -------------------------------------------------------------------------------- Source: based on Armstrong, J.B., Duhon, S.T., and Malacinski, G.M. (1989) Raising the axolotl in captivity. In J.B. Armstrong and G.M. Malacinski (eds.) Developmental Biology of the Axolotl. New York: Oxford University Press, pp. 220-227. Modified protocol (but not recipe) 2/26/98 when we realized that MgSO4 and CaCl2 easily combine and precipitate out. -------------------------------------------------------------------------------- 100% Holtfreter's solution for dejellied embryos, before stage 38 NaCl 3.5 g NaHCO3 0.2 g KCl 0.05 g MgSO4 stock solution 333 µl CaCl2 stock solution 333 µl dH20 1 liter Check pH; should be between 7 and 7.5. 50% Holtfreter's solution for older embryos, larvae, and adults flask carboy trash can NaCl 1.75 g 35 g 262.5 g NaHCO3 0.1 g 2 g 15 g KCl 0.025 g 0.5 g 3.75 g MgSO4 stock solution 335 µl 6.66 ml 50 ml CaCl2 stock solution 335 µl 6.66 ml 50 ml Novaqua 335 µl 6.66 ml 50 ml dH20 1 liter 20 liters 150 liters Check pH; should be between 7 and 7.5. -------------------------------------------------------------------------------- MgSO4 stock solution Combine: MgSO4 300 g deionized water 500 ml CaCl2 stock solution Combine: CaCl2 150 g deionized water 500 ml -------------------------------------------------------------------------------- |
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