In the chart below fill in the genotype of E coli prior to t

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 ? ____

Solution

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.