User:Justin M. Scott/Notebook/Col V Population Dynamics/Entry Base

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Solid Media Experiment 9/18/2012 Placed singles from an LB+CMP plate for BC208pACYC184 (Streaked 9/7/12) 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 #4 (Streaked 9/7/12) 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 9/7/12) in 10mL of LB with 10µL of NAL (1000x) and 20µL of KN (500x) Incubated on roller drum at 37˚C and speed 5 9/19/2012 Spun 1mL of O.N. at 14000 rpm for 3 minutes. Poured off supernatant Resuspended pellet in 1mL of M63 Diluted resuspended pellet to 108 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 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 Incubated at 37˚C for 24 hours Added 300µL 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 Added 200µL 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 Added 100µL 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 Diluted the liquid in the sterile beaker to 107 Used droplet method for viable cell count onto new LB+STR+AMP+CMP, LB+NAL+AMP, LB+NAL+KN plates Incubated at 37˚C overnight Repeated for 72 hours