1 2

HEPES-buffered Modified Steinberg's Solution (HBSt)

(Zackson & Steinberg, 1986, Dev. Biol. 117: 342-353)

NaCl

58.0 mM

3.40 g

KCl

0.67 mM

0.0500 g

CaCl2. 2H2O

0.34 mM

0.0500 g

MgSO4. 7H2O

1.0 mM

0.250 g

HEPES

1.10 g

dH2O

to 1 liter


Adjust to pH 7.4 with NaOH or HCl

Add 140 ml of a 50 mg/ml Gentamycin stock solution to 100 ml 100% HBSt for Operating Solution (70 mg/ml final concentration). Sterile with 0.2 mm filter.

Operating dishes

Operate in 60 mm dishes coated with 1% agarose in 1X HBSt (add 1-2% powdered charcoal for contrast for albino embryos). Right before use heat a glass rod and use to melt several small depressions in the agarose to help confine embryos during surgery.


© Cebra-Thomas 2003
Last Modified: 25 May 2004

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