Gene cloning: fragmentation -> ligation with DNA ligase -> -> insertion into plasmid -> transfection
Gene cloning strategies:
Cloning by expression - a cDNA library is made and transfected into cells such as e.coli. You can detect the successful transfections by looking at the protein the DNA expresses or by radioactively labelling the DNA such that upon autoradiography, they are highlighted.
Cloning by differential expression - comparing the control genetic info to that of a disease. Can be before and after disease event e.g. seizure in epilepsy.
Cloning by homology - degenerate oligonucleotides are made to account for every possible codon. These hybridise with the DNA after it is unwound.
Cloning by protein protein interaction - use knowledge of the protein interaction and its characterisation to determine what it is. If it reacts with the target protein then you have the cDNA that encodes it in the culture. There are 3 main ways of observing protein protein interaction but I wont go into detail, thats for another night. Just know that they are immunofluourescence, GSD Pull Down Assay and Fluourescence Energy Transfer (FRET).