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Transformation of E. coli with pGAL DNA
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(EDVO-Kit 221)
With the newly designed plasmid pGAL, transformed E. coli cells will express the same 2 phenotypic traits as plasmid pUC8: ampicillin resistance (formation of colonies) and a blue color. However, unlike pUC8, pGAL contains the entire lac Z gene for B-galactosidase and inducer IPTG is not required. The blue color is the result of the cleavage of the X-GAL (substrate) by B-galactosidase (enzyme). The lac operon is also discussed. Note: some perishable items require refrigerator or freezer storage. Reagents for 10 lab groups. Duration: transformation – 45 minutes, plating – 5 minutes, incubation – overnight. Note: Some perishable items require refrigeration or freezer storage.
Kit contents:
- E. coli (freeze-dried LyphoCells)
- Plasmid DNA
- Control buffer
- Induction buffer
- Ampicillin
- X-GAL in solvent
- Cell reconstitution media
- Luria broth agar
- Luria recovery broth
- Petri plates
- Transfer micropipets
- 10 mL pipets
- 1 mL pipets
- Inoculating loops
- Microtest tubes
- Instructions
Required, but not included: water baths (to set at 37 and
42°C), incubator/oven (37°C), hot plate or microwave oven, automatic micropipet to deliver 0.5-5 µl, ice, and personal safety equipment.
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