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polymerase chain reaction pcr
polymerase chain reaction
replication of dna
dna replication
dna sequences
How Cloning Works
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Cloning is the process of producing an individual that is the exact genetic copy of another. This means the entire genome of an individual is replicated. Cloning can also involve the replication of DNA sequences. The cloning of organisms can be seen in nature, occuring in the birth of identical twins. However, artificially organism cloning can be accomplished using somatic cell nuclear transfer.

Today, we can obtain, amplify, and manipulate specific DNA fragments using DNA technology. The discovery of thermostable DNA polymerases, such as Taq Polymerase have made it possible to manipulate DNA replication in the laboratory. In an in vivo approach (within live cells), a specific fragment of DNA is inserted into a specially designed plasmid. These plasmids known as vectors, carry and amplify the gene of interest. DNA fragments are separated by an enzyme called restriction endonucleases. Then, the fragmented DNA is inserted into vectors to form recombinant DNA molecules. These recombinant DNA molecules are then transferred into bacterial cells where the molecule is amplified during the division of the bacterial cell. This process called DNA cloning, results in two identical recombinant DNA molecules.

In a similar in vitro approach (in test tube), the polymerase chain reaction (PCR) is used. This method relies on repeated heating and cooling of the reaction for DNA melting and replication of DNA. During this process, a specific gene or DNA region is identified by the binding of specific short primers. These primers serve are the start point for DNA polymersase to come and amplify the DNA. The heating and cooling in this process are necessary as the DNA goes through a process of denaturation, annealing, and elongation. DNA technology is used every day for molecular biology research.

Dolly the sheep was the first mammal ever to be cloned using somatic cell nuclear transfer (SCNT). SCNT involves transferring the nucleus of a somatic cell into an oocyte egg from which the nucleus has been removed. After removing the nucleus, the oocyte can be treated with chemicals to stimulate cell growth and division. If the clones are able to divide normally, it can be transferred into the uterus of the surrogate mother. This process however, does not produce a 100% genetically identical organism. The genetic material in this process comes from the nucleus (the donor DNA) and the mitochondria in the cytoplasm of the enucleated oocyte. A similar method of cloning is called artificial embryo splitting. This is a process in which the embryo is split in the maturation before embryo transfer. If this procedure is done successfully, identical twins are produced.

While certain types of cloning such as asexual reproduction, seen in bacteria, insects, and plants has been taking place for centuries, DNA Replication and animal cloning are relatively new. Scientists have been cloning animals since 1952 beginning with a tadpole. Along with Dolly the sheep, goats, cows, mice, pigs, cats, rabbits, and a gaur have also been created using nuclear transfer technology. The possibility of human cloning exists but ethical reasons have hindered experimentation.


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