Answered: -DNA -DNA/+AMP +DNA/+AMP…

Biology: The Unity and Diversity of Life (MindTap Course List)

14th Edition

ISBN:9781305073951

Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Chapter15: Studying And Manipulating Genomes

Section: Chapter Questions

Problem 12SQ: Which of the following can be used to carry foreign DNA into host cells? Choose all correct answers....

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Overview of Transformation Protocol

-Prepare competent bacteria for transformation:

Treat starter E. coli bacteria with CaCl2and Competent Cell Solution (CCS). Store on ice until transformation procedure.

Competent cells are cells that are likely to take up foreign DNA and be transformed. This step increases the likelihood that the E. coli cells will take up the introduced vector and be transformed.

-Transformation procedure:

Obtain two microcentrifuge tubes containing your competent cells. Label one tube +DNA and one -DNA.

Add CaCl2 to both tubes.

Add the transformation mix containing the plasmid DNA to the tube labeled +DNA. Do not add any plasmid DNA to the -DNA tube.

Incubate both tubes on ice for 10 minutes. Then, place both tubes in a 42\deg C water bath for 45 seconds. Replace the tubes in an ice bucket for 2 minutes.

Add recovery broth to both tubes.

Incubate both tubes in a 37 C water bath for 5 minutes.

Questions:

1)What is the selectable marker in this experiment? How does marker allow for selection of transformed cells only?

2) What is the inducer in this experiment? How will the presence of the inducer influence the gene expression of the transformed cells?

3)On which plate(s) would you expect to find bacteria most like the source E. coli? Why?

4) On which plate(s) would you find only transformed cells? Why?