(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 refriferation 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 0.5–5μl, ice, and personal safety equipment.
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