UNIT 5.9 Deriving Probes From Large-Insert Clones by PCR Methods
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Human Genetics
How to Cite
Albertsen, H., Thliveris, A., Riley, J. H., Munroe, D. J., Watkins, P. and Basson, C. T. 2001. Deriving Probes From Large-Insert Clones by PCR Methods. Current Protocols in Human Genetics. 17:5.9:5.9.1–5.9.37.
- Published Online: 1 MAY 2001
- Published Print: MAY 1998
This unit describes several polymerase chain reaction (PCR)-based methods to obtain DNA fragments from clones with large inserts without prior knowledge of the insert DNA sequence. The protocols can be categorized into three groups: (1) methods to generate DNA fragments at random representing the entire length of the cloned insert, (2) methods to generate DNA fragments representing the extremities of an insert, and (3) methods to generate complex probes suitable for fluorescence in situ hybridization. Support protocols describe direct cloning of these PCR products and the isolation of total yeast DNA from yeast artificial chromosome (YAC) clones.