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


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 liters_________


NaCl_________________110 mM___________________102.85g___________205.71g___________308.55g________
{|border="1" cellpadding="5" cellspacing="0" align="center"
 
|-
Tris base______________15 mM____________________29.075g___________58.15g____________87.23g________
|
 
! scope="col" style="background:#efefef;" |(1x solution)
KCl____________________2 mM____________________2.385g_____________4.77g____________7.2g________
! scope="col" style="background:#efefef;" |
 
! scope="col" style="background:#efefef;" |amounts to make 8X stock
Na2HPO4 (anhydrous)___0.5 mM____________________1.36g______________2.72g____________4.1g________
! scope="col" style="background:#efefef;" |
 
|-
NaHCO3________________2 mM____________________2.69g______________5.38g____________8.07g________
! scope="row" style="background:#efefef;" |
 
|
:Adjust pH to 7.6 with glacial acetic acid (approx. 25 mls for 4 litres)
|2 litres
 
|4 litres
MgSO4.7H2O____________1mM____________________7.89g______________3.945g___________11.84g________
|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"|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.

Revision as of 09:53, 7 December 2011

Protocol submitted by VGP


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

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.

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