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(608) 265-2801/262-9495
Directions For Superovulating Mice
PG600: Purchased through Intervet
405 State St. PO Box 318
Millsboro, DE 19966
800-441-8272
Cat. # PG-720-1 (single dose tank with 5 single doses/box)
$4.20/dose therefore one box of 5 vials =$21.00
1. Add 800µl diluent to 1 dose vial (400 IU PMSG/vial).
2. Aliquot 50µl of PG600 solution to 1.5ml eppendorf tubes, label PG and freeze at -20C. This amount will superovulate 4-5 mice.
Keep hormones frozen until time of use. DO NOT STORE IN A FROST-FREE FREEZER.
3. Before using, thaw the tube of PG-600 (labeled PG). Add 450µl of sterile water to the tube (final volume= 500µl). The final concentration = 5 IU PMSG in each 100µl of solution. One tube is enough for 4 mice.
4. Inject 0.1 cc of PG-600 by IP into each mouse using a 1cc syringe and a 25 gauge X 5/8 inch needle. FVB and C57Bl6 females work best when they are 5-8 weeks of age. PG-600 is usually administered between 4-5 pm (lights off @ 5pm, on @ 5am). The only exception is to collect 1-cell C57Bl6 embryos, PG-600 is administered between noon-1pm.
hCG : hCG (human chorionic gonadotrophin)
Purchased through Organon
West Orange, New Jersey 07052
800-241-8812
10,000 unit vials
1. Add 10ml of solvent to one vial of hCG. Dissolve hormone thoroughly and aliquot 1ml into 1.5ml eppendorf tubes. Label tubes and freeze at -20C. Hormone is now at 1000 units/ml.
2. Transfer 1ml of 1000 unit/ml aliquot to 50ml Falcon tube. Add 19ml filtered sterile saline (0.9% NaCl). Mix and aliquot into 1.5ml eppendorf tubes. We put 500µl/tube which is enough for 4-5 mice. Label tubes with h. Hormone is now at 50 units/ml.
3. 46-48 hrs after PG600 injections, thaw the tube of hCG (labeled h). Use without diluting.
4. Inject 0.1 cc of hCG into each mouse. Mate the female with a male mouse (use a 1:1 ratio of males to females). Check for vaginal plugs the following morning (plug date=day 0.5)
5. One-cell embryos are collected from the oviducts of day 0.5 females. Morula are collected by flushing the oviducts and uteri of day 2.5 females. Blastocysts are collected by flushing the uteri of day 3.5 females.