DNA Cloning

Abstract

Cloning is the process of moving a gene from the chromosome it occurs in naturally to an autonomously replicating vector. In the cloning process, the DNA is removed from cells, manipulations of the DNA are carried out in a test-tube, and the DNA is subsequently put back into cells. Because E. coli is so well characterized, it is usually the cell of choice for manipulating DNA molecules. Once the appropriate combina- tion of vector and cloned DNA or construct has been made in E. coli, the construct can be put into other cell types. This chapter is concerned with the details of the individ- ual steps in the cloning process: 1 How is the DNA removed from the cells? 2 How is the DNA cut into pieces? 3 How are the pieces of DNA put back together? 4 How do we monitor each of these steps?