Final answer:
To isolate and amplify DNA fragments, a positive clone from a genomic library is identified, digested by restriction enzymes, analyzed via gel electrophoresis, and then subcloned into a vector. The cloned DNA with the desired gene is then introduced into a bacterial host for amplification. The numerous copies produced are used for further study or for producing proteins like insulin.
Step-by-step explanation:
Isolating and Amplifying DNA Fragments
To isolate and amplify DNA fragments, and to introduce them into a vector, several steps are involved. Initially, the genome is screened to identify the specific gene of interest. Once a positive clone from the genomic library is identified, it goes through further analyses, such as restriction endonuclease digestion, agarose gel electrophoresis, and Southern blotting. After identification, the gene is subcloned into a suitable vector for propagation. The selected DNA fragment is then amplified, cut from its source, and ligated into a cloning vector, like a plasmid which acts as an independent replicating circular DNA molecule.
Following this, the plasmid containing the desired DNA is introduced into a bacterial host. In this bacterial environment, the recombinant DNA molecule is cloned as the bacterium replicates its own DNA. This results in the production of millions of copies of the cloned DNA over a certain time period, which can then be used for protein production or for further research endeavors.