DNA cloning - Wiley-Blackwell

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Chapter 14DNA cloningCloning is the process of moving a gene from the chromosome it occurs in naturallyto an autonomously replicating vector. In the cloning process, the DNA is removedfrom cells, manipulations of the DNA are carried out in a test-tube, and the DNA issubsequently put back into cells. Because E. coliis so well characterized, it is usuallythe 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 canbe put into other cell types. This chapter is concerned with the details of the individ-ual steps in the cloning process:1How is the DNA removed from the cells?2How is the DNA cut into pieces?3How are the pieces of DNA put back together?4How do we monitor each of these steps?Isolating DNA from cellsPlasmid DNA isolationThe first step in cloning is to isolate a large amount of the vector and chromosomalDNAs.

Chapter 14 DNA cloning 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 …

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