User:Justin M. Scott/Notebook/Col V Population Dynamics/2012/10/09

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Solid Media Experiment 10/9/2012 Methods: Placed singles from an LB+CMP plate for BC208pACYC184 (Streaked 10/5/12) in 10mL of LB with 10µL of CMP (1000x), 10µL of AMP (1000x), and 10µL of STR (1000x) Placed singles from an LB+NAL plate for Exp. Resist #2 (Streaked 10/5/12) in 10mL of LB with 10µL of NAL (1000x) and 10µL of AMP (1000x) Placed singles from an LB+NAL+KN plate for BC212 (Streaked 10/5/12) in 10mL of LB with 10µL of NAL (1000x) and 20µL of KN (500x) Incubated on roller drum at 37˚C and speed 5 10/10/2012 Method: Diluted each O.N. to 107 Pipetted 30µL drops of diluted BC208pACYC184 onto LB plates in an circular patter, 8 drops to a circle Allowed droplet to soak into the media Pipetted 30µL drops of diluted Exp. Resist #5 onto LB plates in an circular patter, 8 drops to a circle Allowed droplet to soak into the media Pipetted 30µL drops of diluted BC212 onto LB plates in an circular patter, 8 drops to a circle Allowed droplet to soak into the media Conducted in triplicate Incubated at 37˚C After 24 hours of incubation, replica plated on LB+STR+AMP+CMP, LB+NAL+AMP, LB+NAL+KN plates and a new LB plate Incubated at 37˚C for 24 hours Replica plated the LB plate onto LB+STR+AMP+CMP, LB+NAL+AMP, LB+NAL+KN plates and a new LB plate Added 1000µL of liquid LB to the LB+STR+AMP+CMP, LB+NAL+AMP, LB+NAL+KN plates Scraped plate with a glass spreader Poured off liquid LB into a sterile beaker on ice Repeated the scrapping twice more (3000µL of liquid LB total for each plate) Diluted the liquid in the sterile beaker to 108 (so a total dilution of 109) Used droplet method for viable cell count onto new LB+STR+AMP+CMP, LB+NAL+AMP, LB+NAL+KN plates Platted droplets in triplicate Incubated all plates at 37˚C overnight Replica plated the LB plate onto LB+STR+AMP+CMP, LB+NAL+AMP, and LB+NAL+KN plates Placed the viable cell count plates in 4˚C refrigerator Scrapped the LB+STR+AMP+CMP, LB+NAL+AMP, and LB+NAL+KN plates Incubated all plates at 37˚C for 24 hours Placed the viable cell count plates in 4˚C refrigerator Scrapped the LB+STR+AMP+CMP, LB+NAL+AMP, and LB+NAL+KN plates Incubated all plates at 37˚C for 24 hours Placed the viable cell count plates in 4˚C refrigerator Counted colonies on viable cell count plates Results: Solid Competition 10.10.12 Notes: Replica platting 1 order: cmp,amp,kn Master 3 fell out during replica platting Replica platting 2 order: amp,kn,cmp Replica 2 fell out during replica platting Replica platting 3 order: kn,cmp,amp