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Holtfreter's Solution




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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.


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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.



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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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