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