Molecular cloning refers to the assembly of DNA molecules into a vector and the subsequent transformation of an organism (often bacteria or yeast). Molecular cloning methods are central to biology and medicine. The term molecular cloning comprises many different techinques. A typical molecular cloning flow looks as follows:
Confirmation of plasmid assembly can be determined by performing DNA sequencing. It is important to confirm that the inserts have been successfully ligated into a plasmid vector with the correct conformation and reading frame. Frame shift can cause nonsense or misense mutations that lead to the expression of nonfunctional proteins.