Description
Advanced – For experienced high school and college classes; requires some technical skill.
Developed in cooperation with the DNA Learning Center, this lab features a deceptively simple experiment that shows the key elements of constructing recombinant DNA. Students ligate (join) DNA fragments containing 2 types of antibiotic-resistant genes and insert the recombined DNA into E. coli cells using rapid colony transformation. Cells containing recombined DNA molecules are detected by their dual antibiotic resistance.
Students use specially developed pBLU® plasmid in a colony transformation procedure to observe the phenotypic effect of inserting new genes into living bacteria. An ampicillin-sensitive strain of E. coli, incapable of producing β-galactosidase for lactose breakdown, is induced to take up pBLU® plasmid DNA. This plasmid contains genes for both ampicillin resistance and β-galactosidase production. The transformed cells are plated on (1) medium containing ampicillin, and (2) medium containing ampicillin and X-gal. (X-gal is a histochemical substrate that, when cleaved by β-galactosidase, forms a blue product.) Transformants appear as white colonies on the ampicillin medium and as blue colonies on the ampicillin/X-gal medium. The transformed colonies’ phenotype is lac+ and ampr.
Kit includes a voucher to request perishables later at your convenience. Contact us or return voucher to request delivery of perishables.
Note: Perishables are shipped on cold gel packs. Foreign orders require special handling.