High-salt solution for egg laying (Zorn lab/Ron Laskey): Difference between revisions
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RAL’s Ca++-free High Salt Barth: Egg Laying (8X) | 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. | |||
Tris | {|border="1" cellpadding="5" cellspacing="0" align="center" | ||
|- | |||
| | |||
! scope="col" style="background:#efefef;" |(1x solution) | |||
Na2HPO4 (anhydrous) | ! colspan="3" style="background:#efefef;" |amounts to make 8X stock | ||
|- | |||
! scope="row" style="background:#efefef;" | | |||
|style="background:#efefef;"| | |||
:Adjust pH to 7.6 with glacial acetic acid (approx. 25 mls for 4 litres) | |style="background:#efefef;"|2 litres | ||
|style="background:#efefef;"|4 litres | |||
MgSO4. | |style="background:#efefef;"|8 litres | ||
|- | |||
! scope="row" style="background:#efefef;" |NaCl | |||
|110 mM | |||
|102.85g | |||
|205.71g | |||
|308.55g | |||
|- | |||
! scope="row" style="background:#efefef;" |Tris base | |||
|15 mM | |||
|29.075g | |||
|58.15g | |||
|87.23g | |||
|- | |||
! scope="row" style="background:#efefef;" |KCl | |||
|2 mM | |||
|2.385g | |||
|4.77g | |||
|7.2g | |||
|- | |||
! scope="row" style="background:#efefef;" |Na2HPO4 (anhydrous) | |||
|0.5 mM | |||
|1.36g | |||
|2.72g | |||
|4.1g | |||
|- | |||
! scope="row" style="background:#efefef;" |NaHCO3 | |||
|0.5 mM | |||
|1.36g | |||
|2.72g | |||
|4.1g | |||
|- | |||
! scope="row" style="background:#efefef;" | | |||
|colspan="4" style="background:#efefef;"|Adjust pH to 7.6 with glacial acetic acid (approx. 25 mls for 4 litres) | |||
|- | |||
! scope="row" style="background:#efefef;" |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. | Make 8X stock, dilute to 1x solution in dH2O just before use. Maximum recommended storage: 8 weeks at 4oC. |
Latest revision as of 10:13, 7 December 2011
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.