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Despite this, the low efficiency of the technique has prompted some researchers, notably Ian Wilmut, creator of Dolly the cloned sheep, to abandon it.
Tools and reagents
Nuclear transfer is a delicate process that is a major hurdle in the development of cloning technology. Materials used in this procedure are a microscope, a holding pipette (small vacuum) to keep the oocyte in place, and a micropipette (hair-thin needle) capable of extracting the nucleus of a cell using a vacuum. For some species, such as mouse, a drill is used to pierce the outer layers of the oocyte.
Various chemical reagents are used to increase cloning efficiency. Microtubule inhibitors, such as nocodazole, are used to arrest the oocyte in M phase, during which its nuclear membrane is dissolved. Chemicals are also used to stimulate oocyte activation. When applied the membrane is completely dissolved.
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Somatic cell nuclear transfer
Somatic Cell Nuclear Transfer (SCNT) is the process by which the nucleus of an oocyte (egg cell) is removed and is replaced with the nucleus of a somatic (body) cell (examples include skin, heart, or nerve cell). The two entities fuse to become one and factors in the oocyte cause the somatic nucleus to reprogram to a pluripotent state. The cell contains genetic information identical to the donated somatic cell. After stimulating this cell to begin dividing, in the proper conditions an embryo will develop. Stem cells can be extracted 5–6 days later and used for research.
Reprogramming
Genomic reprogramming is the key biological process behind nuclear transfer. Currently unidentified reprogramming factors present in oocytes are capable of initiating a cascade of events that can reset the mature, specialized cell back to an undifferentiated, embryonic state. These factors are thought to be mainly proteins of the nucleus.