02 - Code Breakers: Using CRISPR to Rewrite Genetics
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Cat. #221 Transformation of E.coli with pGAL™ (Blue Colony) For 10 groups. In this experiment, your students can see a blue color change in transformed cells due to the switching on of a gene. The pGAL™ plasmid gives them a blue color due to the production of the ß-galactosidase protein by the lacZ gene. IPTG is not required in this experiment since pGAL™ contains the complete lacZ gene. Cat. #222 Transformation of E.coli with Blue and Green Fluorescent Proteins For 10 groups. Green Fluorescent Protein (GFP), which is responsible for bio- luminescence in the jellyfish Aequorea victoria , is used extensively in all areas of science. Many organisms have been transformed with the GFP gene. It has proven to be so useful that scientists have mutated it to produce Blue Fluo- rescent Protein (BFP). In this simple experiment, your students will transform bacteria either with GFP, BFP or both! Cat. #223-AP08 Transformation of E.coli with Green Fluorescent Proteins For 10 groups. Transformed cells take up a plasmid containing the GFP gene. The GFP gene was isolated from the jellyfish Aequorea victoria . Transformed colonies expressing the GFP protein are visibly green in normal light but will fluoresce brightly when exposed to long wave UV light. Cat. #300 Blue/White Cloning of a DNA Fragment & Assay of ß-galactosidase For 5 groups. When DNA is subcloned in the pUC polylinker region, ß-galacto- sidase production is interrupted, resulting in the inability of cells to hydrolyze X-Gal. This results in the production of white colonies amongst a background of blue colonies. This experiment provides a DNA fragment, linearized plas- mid, and T4 DNA Ligase. Following the ligation to synthesize the recombinant plasmid, competent E.coli cells are transformed and the number of recombi- nant antibiotic resistant white and blue colonies are counted. ß-galactosidase activity is assayed from blue and white bacterial cells. This experiment can be broken down into three modules: ligation, transformation, and assay of ß-galactosidase. Cat. #301 Construction & Cloning of a DNA Recombinant For 5 Plasmid Constructs & Analyses. Cloning is frequently performed to study gene structure, function, and to enhance gene expression. This experiment is divided into five modules. Clones are constructed by ligation of a vector and a fragment insert. The constructs are then transformed into competent cells and the cells are grown and selected for resistance. Plasmid DNA is then isolated from the transformants, cleaved with restriction enzymes, and analyzed by agarose gel electrophoresis. Recommended for college level courses.
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