UNIT 3.16 Subcloning of DNA Fragments
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Struhl, K. 2001. Subcloning of DNA Fragments. Current Protocols in Molecular Biology. 13:IV:3.16:3.16.1–3.16.2.
- Published Online: 1 MAY 2001
- Published Print: JAN 1991
The essence of recombinant DNA technology is the joining of two or more separate segments of DNA to generate a single DNA molecule that is capable of autonomous replication in a given host. The simplest constructions of hybrid DNA molecules involve the cloning of insert sequences into plasmid or bacteriophage cloning vectors. The insert sequences can derive from essentially any organism, and they may be isolated directly from the genome, from mRNA, or from previously cloned DNA segments (in which case, the procedure is termed subcloning). Alternatively, insert DNAs can be created directly by DNAsynthesis. This unit provides protocols for the subcloning of DNA fragments and ligation of DNA fragments in gels.