Veterinary Andrology and Reproductive Techniques

BUFFER

• Prepared under separate laminar air flow unit.
• Use autoclaved triple distilled water or Milli-Q water for buffer preparation.
• Use only GR or AR grade chemicals.
• Autoclave the buffer at 5 psi for 20 minutes.
• After autoclaving bring the buffer to room temperature before diluent preparation.

• The buffer can be prepared one day before preferably in the previous day evening of the collection.
• The chemicals used for buffer preparation should be Graded reagent (GR) or Analytical grade (AR) grade and should be purchased from standard companies.
• Laboratory grade (LR) chemicals should not be used due to low purity.
• Whenever new chemicals are purchased it is recommended to conduct the post thaw survival rate by conducting few split ejaculate trials (maintaining the previous batch as control) with the new chemical.
• Buffer should be prepated in a sterile manner in the Buffer and Diluent preparation room.
• Buffer should be sterilized by autoclaving at 5 psi pressure of 20 minutes.
• Upon cooling and after addition of egg yolk, antibiotics should be added under laminar flow unit.
• The buffer should not be stored and reused in subsequent days.