Enucleation Pre-Enucleation:

Oocytes (unfertilized embryos) are incubated with a DNA specific stain to identify all DNA material that needs to be removed.

Enucleation:

A pipette holds the oocyte while a needle is inserted through the zona pellucida of the oocyte. The Chromosomes and the polar body are drawn into the needle removing this material from the oocyte.

Enucleation Complete:

Once this DNA material is removed, all that remains inside the zona pellucida is the cytoplasm. The oocyte is ready to receive the DNA material of the animal being cloned.

Transfer of DNA

Fibroblast pickup:

A fibroblast cell is selected from the founder’s cell line and loaded into the needle.

DNA Transfer:

The needle loaded with the fibroblast cell is injected into the hole in the zona. The cell is placed between the inner wall of the zona pellucida and the cytoplasm.

Transfer complete:

When completed the result is a nuclear transfer couple where the fibroblast cell is separate from the oocyte cytoplasm.

Fusion
Fusion 1:

The transferred oocytes are exposed to an electrical stimulus that fuses the fibroblast cell into the oocyte cytoplasm.

Fusion 2:

The fibroblast cell’s nucleus, with the founder animal’s complete set of genes, enters the oocyte’s cytoplasm. Cellular development can be seen within the embryo in about 20 hours after fusion

Embryo development and Transfer of Embryo:

The embryos are then grown in incubators for seven days. At this time the embryos are evaluated and the high quality embryos are ready for transfer.

Clone Embryo Transfer:

Once the NT or cloned embryos have been evaluated they are transferred into recipients or host donors.

The Birth

Nine months later, the clone calf is born under special care of a team of experts.

The Road Home:

60 to 90 days after birth the cloned calves have passed their physical exams and are ready to make the trip to their owner’s farm or ranch.