In the chart below, fill in the genotype of E. coli prior to transformation and

Question

In the chart below, fill in the genotype of E. coli prior to transformation and after transformation. What was the purpose of transferring the +DNA and -DNA tubes from ice, to hot water, to ice again? ____ Why were the vials incubated for 10 minutes in LB broth rather than transferring their contents directly to the plates? ____ What information is provided by the LB/-DNA plate? ____ Obviously, transformation could only occur if the pGLO plasmid was introduced into the solution. Which plate(s) exhibit transformation? ____ What information is provided by the LB/amp/-DNA plate? ____ Why does the LB/amp/ara/+DNA plate fluoresce when the LB/amp/+DNA plate does not ? ____

Explanation / Answer

1. Before transformationin 1-no GFP, after GFP+; bla-no, after bla+

2. Heat shock- rapid change in the temperature makes the Ecoli cell walls permeable to plasmid DNA.
3. 10 minutes in LB Broth gives the transformed cells time to grow-immediately begins to produce ampicillin resistance gene.
4. Control- verifies that Ecoli can grow in LB media without ampicillin.
5. +DNA/LB/AMP;+DNA/LB/AMP/ARA
6. COntrol-verifies that Ecoli can grow in media containing ampicillin.
7.Green Fluorescence Protein is expressed only in the presence of Arabinose (inducer), turns on the operon.

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