November
23
NOVEMBER 16TH and 19TH- LABS
- NOVEMBER 16TH, 2018
- OBJECTIVE:
- To pick a plaque and create serial dilutions, so dilutions can be easily plated next time coming into lab
- PROCEDURE:
- Tables were cleaned and lamps were lit
- Seven micro-centrifuge tubes were labeled according to the dilution (10^0 through 10^-6)
- The tube labeled 10^0 was filled with 100𝝁L of phage buffer and tubes 10^-1 through 10^-6 were filled with 90𝝁L of phage buffer
- The P4 plate was examined under a light microscope, where a plaque was picked using a micropipette
- The pipet tip was then swirled in 100𝝁L of phage buffer
- Then 10𝝁L of the 10^0 dilution was pipetted into the 10^-1 tube, then 10𝝁L of solution from the 10^-1 tube was pipetted into the 10^-2 tube, and so on until a dilution of 10^-6 was reached
- After the plaque was picked 8mL of phage buffer was put onto the P4 plate, where the plate was then left to flood for 48 hours
- RESULTS:
- None
- CONCLUSION:
- None
- NEXT STEPS:
- Plate dilutions and filter lysate from plate flooding
- OBJECTIVE:
- NOVEMBER 19TH, 2018
- OBJECTIVE:
- To plate serial dilutions and filter lysate
- PROCEDURE:
- Tables were cleaned and lamps were lit
- A large test tube was filled with:
- 4mL LB
- 5mL 2X TA
- 45𝝁L of CaCl2
- 4.5mL was then pipetted onto 2 plates, one plate where the serial dilutions will be tested, and the control
- After allowing 10 minutes for the plates to solidify, 10𝝁L of each dilution was pipetted onto the designated location on the plate
- After allowing 10 minutes to let the dilutions sit on the plate, the plate was placed right side up in the incubator
- The contents of the flooded plate were poured into a top filter, and filtered out creating a new lysate
- RESULTS:
- The results can be viewed in Figure 25
- CONCLUSION:
- The area on the plate labeled 10^-3 will be used as the high titer for the lysate
- NEXT STEPS:
- Calculate how much is needed to make a high titer, then plate the amount calculated
- Calculate how much is needed to make a high titer, then plate the amount calculated
- OBJECTIVE:
