1.Preparation of extender : Add powder to provided double distilled, sterile water. ( Mix well-until powder is completely dissolved.) Add 20ml fresh egg yolk. (See seperate description of methodology to prepare egg yolk). 2. Collect sperm-rich fraction. 3. Exend in Canine-EXT mixture at room temmperature. If sperm concentration of raw ejaculate is 300-500 x10/ml then dilute 1 part semen & 3parts extender. If sperm concentration of raw ejaculate is 500-800 x 10/ml then dilute 1 part semen & 4 parts extender. If sperm concentration of raw ejaculate is over 800 x 10/ml then dilute 1 part semen & 5 part extender. 4. Cool for 60-75 minutes to 4-5 degrees Celcius. This can best be done by tightly sealing a tube of extended semen and then fill a beaker with room temperature water(volume of water should be approximately 10x that of extended semen in tube) and then place tube of extended semen in beaker with water. Place beaker in 4-5 degrees celcius refrigerator and allow to cool slowly.
Preparation of Fresh Egg Yolk for use in semen extender:
1. Obtain fresh large eggs. Aquire from a supplier with a fast turn-over rate. Eggs on the store shelf may not be fresh...the fresher the better!
2. Carefully crack eggs onto a clean piece of freezer paper or other suitable surface such as a clean paper towel(absorbent works well) This must be done in a manner that the yolk remains intact.
3. Using an all plastic (10-20ml) syringe, carefully insert the tip of the syringe into the yolk. Withdraw as much of the yolk as possible without contaminating the yolk with egg white or membranes.
CAUTION: Egg whites are damaging/toxic to sperm! Any contaminated yolk must be discarded! Do not use yolks contaminated with blood. Prepare yolk on a daily basis.
Extender, Canine EXT - Chilled Semen....