Transformation Lab Quiz Review

pGLO Plasmid

The small circular piece of DNA inserted into E. coli during transformation.

LB (Lysogeny Broth)

The nutrient-rich media used to provide food for E. coli growth.

Ampicillin

The antibiotic used to select for successfully transformed bacteria.

Ampicillin Mechanism of Action

Inhibition of cell wall synthesis.

bla Gene Product

Beta-lactamase, an enzyme that breaks down ampicillin.

bla Gene

The gene that codes for beta-lactamase to provide ampicillin resistance.

araC Gene

The regulator gene that controls transcription of the GFP gene.

pBAD

The promoter on the pGLO plasmid where RNA polymerase binds to transcribe GFP.

GFP Gene

The reporter gene that codes for Green Fluorescent Protein under UV light.

Arabinose

The sugar that must be present to trigger GFP expression.

Transformation Solution

Calcium chloride (CaCl₂), used to prepare cells for transformation.

Permeability Step (Heat Shock)

Placing cells in a 42°C hot water bath to allow plasmid entry.

Transformation

The process where a bacterium absorbs DNA from its external environment.

Ice Incubation (Post-Heat Shock)

Placing cells on ice to close pores and retain the plasmid.

LB Plate Growth

Both transformed (+DNA) and non-transformed (-DNA) E. coli will grow.

LB/Amp Plate Growth

Only transformed (+DNA) E. coli will grow.

Fluorescence Requirements

LB, Ampicillin, Arabinose, and +DNA cells.

Competent Cells

Bacteria that have the ability to take up foreign DNA from their environment.

Still learning (2)

You've started learning these terms. Keep it up!