UNIT 6.4 Using Synthetic Oligonucleotides as Probes
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Duby, A., Jacobs, K. A. and Celeste, A. 2001. Using Synthetic Oligonucleotides as Probes. Current Protocols in Molecular Biology. 2:II:6.4:6.4.1–6.4.10.
- Published Online: 1 MAY 2001
- Published Print: JAN 1990
The protocols in this unit describe procedures for using mixtures of 32P-labeled oligonucleotides to screen recombinant DNA clones bound to nitrocellulose filters. A partial amino acid sequence of a protein is used to predict the nucleotide sequence of the gene that would encode it. A mixture of oligonucleotides is chosen that includes all possible nucleotide sequences encoding that amino acid sequence. This mixture of oligonucleotides is then used to screen a recombinant DNA library for the corresponding clones. In some cases however, the exact nucleotide sequence of a desired clone is known and it is possible to use a unique oligonucleotide as a probe.