High-salt solution for egg laying (Zorn lab/Ron Laskey)

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Protocol submitted by VGP

RAL’s Ca++-free High Salt Barth: Egg Laying (8X) (Ron Laskey)

Place ovulating females in 1X high salt buffer, 3 liters per female.

Eggs laid in the high salt buffer remain viable for fertilization and microinjection.

(1x solution) amounts to make 8X stock
2 litres 4 litres 8 litres
NaCl 110 mM 102.85g 205.71g 308.55g
Tris base 15 mM 29.075g 58.15g 87.23g
KCl 2 mM 2.385g 4.77g 7.2g
Na2HPO4 (anhydrous) 0.5 mM 1.36g 2.72g 4.1g
NaHCO3 0.5 mM 1.36g 2.72g 4.1g
Adjust pH to 7.6 with glacial acetic acid (approx. 25 mls for 4 litres)
MgSO4.7H2O 1mM 7.89g 3.945g 11.84g

Make 8X stock, dilute to 1x solution in dH2O just before use. Maximum recommended storage: 8 weeks at 4oC.