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The inverted plates were incubated in the 37 o C incubator overnight (24-48 hours).

Using the F250 micropipette, 250 µ l of CaCl 2 solution was added to both the ‘-DNA’ and ‘+DNA’ microcentrifuge tubes.

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The inverted plates were incubated in the 37 o C incubator overnight (24-48 hours).

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  1. Using the F250 micropipette, 250 µl of CaCl2 solution was added to both the ‘-DNA’ and ‘+DNA’ microcentrifuge tubes.

  2. Using a sterile inoculating loop, Escherichia coli colonies were selected from the agar plate and transferred to the ‘-DNA’ and ‘+DNA’ microcentrifuge tubes by twisting the loop vigorously in the CaCl2 solution. The bacterial clumps were broken up using either the vortex or the knuckle boxing method.

  3. The microcentrifuge tubes were incubated at 42oC for 90 seconds in the dry bath, and then immediately returned to the ice rack for a 2 minute incubation period.

  4. The Escherichia coli cells from the ‘-DNA’ and ‘+DNA’ tubes were plated onto the ampicillin-containing agar plates labeled Amp/-DNA and Amp/+DNA respectively. The cells were spread on the plates using a sterile inoculating loop.

  5. The inverted plates were incubated in the 37oC incubator overnight (24-48 hours). Check out the results to see how successful your group was at transforming bacteria

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