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Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. [1] The use of the word cloning refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules.
The choice of vector for molecular cloning depends on the choice of host organism, the size of the DNA to be cloned, and whether and how the foreign DNA is to be expressed. [7] The DNA segments can be combined by using a variety of methods, such as restriction enzyme/ligase cloning or Gibson assembly. [citation needed]
Another example of artificial cloning is molecular cloning, a technique in molecular biology in which a single living cell is used to clone a large population of cells that contain identical DNA molecules. In bioethics, there are a variety of ethical positions regarding the practice and possibilities of cloning.
Recombinant DNA (rDNA), or molecular cloning, is the process by which a single gene, or segment of DNA, is isolated and amplified. Recombinant DNA is also known as in vitro recombination . A cloning vector is a DNA molecule that carries foreign DNA into a host cell , where it replicates, producing many copies of itself along with the foreign DNA.
An example is lacZ-α which codes for the N-terminus fragment of β-galactosidase, an enzyme that digests galactose. A multiple cloning site is located within lacZ-α, and an insert successfully ligated into the vector will
A plasmid is a double stranded circular DNA molecule commonly used for molecular cloning. Plasmids are generally 2 to 4 kilobase-pairs (kb) in length and are capable of carrying inserts up to 15kb. Plasmids contain an origin of replication allowing them to replicate inside a bacterium independently of the host chromosome.
Another method of cloning without the use of DNA digest and ligase is by DNA recombination, for example as used in the Gateway cloning system. [3] [4] The gene, once cloned into the cloning vector (called entry clone in this method), may be conveniently introduced into a variety of expression vectors by recombination. [5]
Gibson assembly is a molecular cloning method that allows for the joining of multiple DNA fragments in a single, isothermal reaction. It is named after its creator, Daniel G. Gibson, who is the chief technology officer and co-founder of the synthetic biology company, Telesis Bio.