Some transgenic animals may prove to be infertile through an inability to mate or rear a litter. It will therefore be necessary to perform an in vitro fertilization (IVF) procedure (I) in order to continue the line. The technique involves superovulating females and fertilizing the eggs with sperm taken from the dissected epididymis of males. IVF eggs are then transferred to a 0.5-d pc pseudopregnant recipient mother. The animals used will depend on the particular experiment; if C57BL/ 6J x CBA/J F1 hybrids were used for the original transgenic production, these should also be chosen for the IVF procedure. Males should be at least 6 wk of age and preferably studs that were rested on the preceding night. Females should be at least 4 wk of age. The described protocol employs a single male and 10 females.
Was this article helpful?