October
29
October 29 2018 Serial Dilutions- Soil C
Rationale: The purpose of this lab is to perform serial dilutions.
Description of Procedures:
- The workstation was cleaned using aseptic technique and an ethanol burner was lit to create an aseptic zone.
- 90 ul of phage buffer was added to four tubes.
- 10 ul of phage buffer lysate from the picked plaque was added to to the 10^-1 tube. 10 ul of the 10^-1 buffer was then added to the 10^-2 tube. This was done out to a dilution of 10^-4.
- 10 ul of each tube was added to 0.5 ml of arthrobacter and allowed to sit for 10 minutes.
- 10 ml of LB broth and 112.5 ul of CaCl2 were added to a tube. 12.5 ml of 2x TA were added to the tube.
- 4.5 ml of the top agar solution was added to the 0.5 ml of arthrobacter and poured onto plates labeled LIP 10-29-18 PA 10^-1, LIP 10-29-18 PA 10^-2, LIP 10-29-18 PA 10^-3, and LIP 10-19-18 PA 10^-4. The rest of the top agar solution was poured onto a plate labeled LIP 10-29-18 control.
- The plates were allowed to sit for 10 minutes and then inverted and stored in the incubator until the next lab.
- The workstation was cleaned using aseptic technique and materials were properly stored and disposed of.
Observations:
- Bottom top agar had not set correctly on some plates.
- Bubbles seen on plates.
Interpretations/Next Steps:
The procedure was complete. The next step will be to make a webbed plate.