Prior to Lab Day:

1. Review the attached lab procedure PDF to familiarize yourself with the technique

Read pages 5-9 for background information on HOW the experiment works.

Use page 10 to formulate your hypothesis and come up with your experimental design.

2. View the attached powerpoint and answer the 4 review questions at the end as a "pre-lab" section of your formal report.

3. Read below...

Lab Day:

1. You will work in your assigned lab groups.

STUDENTS WHO HAVE ALLERGIES TO ANTIBIOTICS SHOULD NOT PARTICIPATE IN THIS LAB

allergic or absent, you should do this: go to https://www.dnalc.org/harlemdnalab/BacterialTrans.html, use the information and simulations to write a hypothetical lab complete with introduction, thorough background, materials, procedure, and predicted outcome with reasoning. Conclude with a section explaining the importance, significance, and application of transformation techniques in science.

2. Before you begin, safety first...

Gloves, goggles, and aprons, and closed-toed shoes MUST be worn for this experiment.

NO MOUTH PIPETTING EVER.

The E. coli bacteria used in this experiment is not considered pathogenic, however good practice will be followed in the handling of said bacteria.

NO food or drink or gum anywhere near the lab tables. At all.

Wipe down lab tables with disinfectant before and after experiment.

Dispose of all materials properly in the appropriate bag marked for biohazard.

WASH HANDS BEFORE LEAVING THE ROOM.

3. Execute the procedure TO THE LETTER on pages 13-16 (steps 1-22) on day 1. Steps 23-24 on day 2.

4. Clean up.

5. When finished, immediately begin writing up a formal lab report in your actual lab notebook of what you did.

a. Title and date

b. Problem: How does the process of bacterial transformation work?

c. Hypothesis: Base it on the components of the transformation system. See pages 10-11 of the packet for help.

ex: If the bacteria are successfully transformed with the plasmid DNA, then those cells will be selected for in the presence of... and will express....

d. Experimental Design: use the brief description and overview on pages 10 and 11 as a guide.

e. Materials: list the items used in the lab - include physical equipment and reagents

f. Procedure: Give a summary of the procedure using pages 13-16 of the lab.

g. Data: draw or photograph your observations for each plate upon viewing after incubation.

For each of the plates record how much bacterial growth you observe (lawn or colonies - determine a count), and the color of the growth.

If you did not get any growth, what factors could be attributed to this fact?

h. Analysis: Determine transformation efficiency using the equation on page 19 of the handout.

Include statements of what you see on the various plates.

i. Conclusion: Did your experiment work. Why/how did it work? or why not?

Answer the 4 questions (in paragraph form) on page 17 as part of your conclusion/discussion section, making sure to EXPLAIN your thoughts.

j. Post-lab: Answer the study questions on page 20 of the lab.

6. Turn your in lab notebook by the assigned date for review by Dr. Kostenko.