Investigate Transformation, a Basic Principle of Genetic Engineering
- Genetically engineer non-pathogenic bacteria
- Materials for 5 lab groups
- Time Required: 90 minutes
In this experiment, the bacteria E. coli JM 101, which is sensitive to ampicillin and incapable of producing beta-galactosidase needed for lactose breakdown, is transformed by insertion of the pUC8 plasmid, receiving antibiotic resistance as well as the lac+ phenotype. In two 45-minute lab sessions, the traits are transferred, and the transformed cells are grown on agar plates containing the antibiotic and on plates with ampicillin and x-gal, a histochemical substrate for β; -galactosidase that yields a blue precipitate upon hydrolysis. Transformants appear as white colonies on plates with ampicillin and blue colonies on plates with ampicillin and x-gal. It comes with enough materials for five setups, as well as a teacher’s guide and student copymaster.
Materials required, not included: incubator and a microcentrifuge
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