1. Medical Interventions Hewitt-Trussville High School September 28, 2011 Bacterial Conjugation Lab

2. Objective Discuss the following: Bacterial conjugation Antibiotic resistance Lab Procedure Streaking Aseptic Technique Perform antibiotic resistance confirmation Predict results of resistance

3. Pneumococcus Bacteria found in the back of the nose of a healthy person When it moves, it can cause ear infection, sinus infection, pneumonia, and meningitis Before 1940, a pneumococcus infection was deadly. What changed? What has this done to the bacteria over time? http://textbookofbacteriology.net/S.pneumoni ae.html

4. Genetic Recombination of Bacteria Transformation Transduction Conjugation http://microculture.tumblr.com/post/6293909843 /fyeahmedicine-bacterial-conjugation-method-of

5. Conjugation Plasmid Extra chromosomal DNA Double stranded and circular Contain from 3 to 25 genes F (fertility) factor Found in E. Coli Regulates formation of pili http://biology.kenyon.edu/courses/bio l09/plasmid/plasmidtut7.htm

6. Antibiotic Resistance

7. Prep Work Melted LB agar LB agar = Lysogeny broth Nutrient rich medium for bacteria to grow Added antibiotics to melted agar First = added streptomycin (LB + str) Second = added ampicillin (LB + amp) Third = added both antibiotics (LB + str + amp) Created the E. coli broths from the slant Strain I = Streptomycin Resistant Strain II = Ampicillin Resistant

8. Directions Wipe down the desk with the cleaning wipes Collect four agar plates LB LB + str LB + amp LB + str + amp Write your initials on the plate Draw a line down the middle of the bottom side of the plate Label one side as I Label the other side as II Do this for all 4 plates

9. Aseptic Technique http://www.youtube.com/watch?v=tBmNitxvqyc&safety_ mode=true&persist_safety_mode=1

10. Directions Place the loop with the Strain I directly under where you labeled the I Streak the plate as shown. http://faculty.ccbcmd.edu/courses/bio141/labmanua/lab2/st reak_an.html http://faculty.ccbcmd.edu/courses/bio141/labmanua/lab2/st reak_an.html Complete these same steps for Strain II on the other side of the plate. Complete this for all four plates.

11. III

12. I

13. I

14. Directions Allow the liquid to soak into the agar. INVERT the plates and place in a stack of four in the incubator In the prep room Wipe all of the desk down with the wipes Wash hands Complete the chart. ***Place all used inoculation loops into the biohazard bag in the prep room DO NOT THROW THESE AWAY!

15. Data Chart Agar PlatesE. Coli Strain I Predicted ResultsE. Coli Strain II Predicted Results LB only LB + str LB + amp LB + str + amp

16. Materials 4 Disposable inoculation loops Aprons Goggles Gloves Bunsen Burner 4 Agar plates (LB, LB+str, LB+amp, LB+str+amp) 2 E. Coli broths (Strain I and Strain II)