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Restriction enzyme mapping of bacterial urease genes: using degenerate primers to expand experimental outcomes
Author(s) -
Reed Kelynne E.
Publication year - 2001
Publication title -
biochemistry and molecular biology education
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.34
H-Index - 39
eISSN - 1539-3429
pISSN - 1470-8175
DOI - 10.1111/j.1539-3429.2001.tb00131.x
Subject(s) - urease , restriction enzyme , polymerase chain reaction , agarose gel electrophoresis , agarose , gene , bacteria , biology , southern blot , enzyme , microbiology and biotechnology , restriction map , computational biology , genetics , biochemistry , plasmid
Two degenerate primers are described that can be used to amplify a 340 bp 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. © 2001 IUBMB. Published by Elsevier Science Ltd. All rights reserved.