Preparation of the Estadiol Solution
- Keep track of the amount of ETOH you use to dissolve
the estradiol to add to your control solution.
- Do NOT use deionized or distilled water!!! That will
kill the fish! A recipe for artificial spring water is below. Otherwise,
dechlorinated tap water is fine.
- Dissolve 2.74 mg in 1 ml ETOH to make a 10 mM solution
of estradiol. This is required because estradiol has low solubility in water.
This is the stock solution.
- Take 220 uL of the 10 mM stock solution into 10
mL of H2O to make a 220 um solution (add ETOH if it doesn't
completely dissolve).
- Add 1 mL of the 220 um solution to 1 L of water
for a final concentration of 60 ug/L estradiol. This is twice the
concentration of estradiol in a typical birth control pill.
- Dose the fish in the liter of water for 3-4 days before
the laboratory. The fish should be kept in the estradiol solution for the
remainder of the experiment. This solution is good for at least 1.5 weeks or
so.
Preparation of Holtfreter’s solution (artificial spring
water)
All Done!