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|
|Tris base||15 mM||29.075g||58.15g||87.23g|
|Na2HPO4 (anhydrous)||0.5 mM||1.36g||2.72g||4.1g|
|Adjust pH to 7.6 with glacial acetic acid (approx. 25 mls for 4 litres)|
Make 8X stock, dilute to 1x solution in dH2O just before use. Maximum recommended storage: 8 weeks at 4oC.