UW Transgenic Animal Facility

(608) 265-2801/262-9495

  Directions For Superovulating Mice

Keep hormones frozen until time of use.  DO NOT STORE IN A FROST-FREE -20°C FREEZER.

PMSG (Pregnant Mare Serum Gonadotropin): Purchased through National Hormone and   

Peptide Program-UCLA         

• Hormones are in 1ml and 500µl aliquots.  1ml will superovulate 8-9 females; 500µl will superovulate 4 females.

• Before using, thaw the tube of PMSG (labeled P). The final concentration = 5 IU PMSG in each 100ul of solution.

• Inject  0.1 cc of PMSG 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 21-27 days of age (10-15 grams).  PMSG is usually administered between 3-5 pm (lights off @ 5pm, on @ 5am.

hCG: (human chorionic gonadotrophin):  Purchased through Sigma

• Hormones are in 500µl aliquots.  This is enough for 3-4 mice.

• 46-48 hrs after PMSG injections, thaw the tube of hCG (labeled h). 

• Inject 0.1 cc of hCG by IP 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)

• Two-cell embryos are collected in M2 from the oviducts of day 1.5 females.  Morula are collected by flushing the oviducts and uteri of day 2.5 females.

• Collect reproductive tracts from both females “with plugs” and “without plugs”, but keep the two groups separated and labeled accordingly.  Bring the tracts over to room 3220 Genetics/Biotech and deliver to Ryan or Kathy.