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Biochemistry and Molecular Biology Education, Vol. 29 (6) (2001) pp. 239-244
© 2001 IUBMB. Published by Elsevier Science Ltd. All rights reserved.
PII: S1470-8175(01)00095-9

Laboratory exercise

Restriction enzyme mapping of bacterial urease genes: using degenerate primers to expand experimental outcomes

Kelynne E. Reed * kreed@austinc.edu

Department of Biology, Austin College, Sherman, Austin, TX 75090, USA

Abstract

Two degenerate primers are described that can be used to amplify a 340bp fragment of the ureC gene from a variety of urease producing bacteria. A series of experiments are outlined that enable students to develop restriction maps of the urease gene clusters from different bacteria. Students learn a variety of molecular biology techniques including polymerase chain reaction, agarose gel electrophoresis, and Southern blotting. This project allows for multiple experimental outcomes using the same two PCR primers.

Keywords: Urease; PCR; Southern hybridization; Restriction mapping

*Tel.: +1-903-813-2064; fax: +1-903-813-2420

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