The experiments presented in this workshop are part of a semester-long inquiry-based molecular biology laboratory course offered to fourth year biological science undergraduate students. The experiments presented here can however be easily adapted to second or third year level laboratories. In the lab course, E. coli transposon mutants were created using a suicide plasposon (a plasmid containing a transposon). The transposon mutants were then screened for auxotroph mutants and the auxotrophs obtained were further characterized phenotypically and genotypically. This workshop focuses on the isolation and phenotypic characterization of auxotroph mutants. During the workshop, the participants will be introduced to the use of a plasposon, pRL27 (containing the Tn5-RL27 transposon), to create random transposon mutation. Participants will then screen for auxotroph mutants, and identify and confirm the phenotype of the mutants using plating techniques. Analysis of the results and ability to adapt the experiment for the screening of other types of mutants will be discussed.
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