GENETIC ENGINEERING & TRANSFORMATION
2024-2025 EDVOTEK ® RESOURCE GUIDE
Cat. #301 Construction & Cloning of a DNA Recombinant
For 5 Plasmid Constructs & Analyses. Cloning is frequently performed to study gene structure and function, and to enhance gene expression. This experiment is di- vided into five modules. Clones are constructed by ligation of a vector and a frag - ment 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 electro- phoresis. Recommended for college-level courses.
Module I: Ligation 70 min.
Electrophoresis 45 min. Module II: Transformation 70 min. Overnight incubation Module III: Culture 15 min. Module IV: Extraction 65-80 min. Module V: Restriction Enzyme Reaction 70 min.
Storage: Some Components Require Refrigerator and Freezer Storage Upon Receipt. CLICK HERE For Experiment #301 Components and Requirements.
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-galactosidase 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 together with a linear plasmid and T4 DNA Ligase. Following the ligation to synthesize the recombinant plasmid, competent E. coli cells are transformed and the number of recombinant 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. Recommended for college-level courses.
Module I: Ligation 70 min. Module II: Transformation and Selection 60 min. Overnight incubation Module III: Assay of ß-galactosidase 60 min.
Storage: Some Components Require Refrigerator and Freezer Storage Upon Receipt. CLICK HERE For Experiment #300 Components and Requirements.
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