Title: Flooded Plate

Date: 31 October 2018

Rationale: The plate that contained 20 microliters of lysate webbed, while the plate containing 30 microliters didn’t (due to the top agar not setting correctly). The lysed plate will be flooded in order to collect phage and, in order ot determine titer, a plaque assay will be done and the plaques counted.

Procedure: Under an aseptic zone

• 4 mL of phage buffer was poured onto the plate and placed onto a plate shaker for 1.5 hours to dislodge phage
• After 1.5 hours, a plaque assay was done to determine titer
  • the following recipe was used for 2 plaque assays:
    • 4 mL LB Broth
    • 5 mL 2x Top Agar
    • 45 microliters CaCl2

~4.5 mL pipetted into a tube containing 0.5 mL Arhtrobacter and either 10 or 20 microliters of lysate. The plates were cooled for 15 minutes and incubated.

Conclusions: After growing both of these plaque assays, the titer will be calculated and hopefully a high titer will result. If not, the plate webbing process will have to be revisited and other methods investigated to discover the block from getting a high titer. Perhaps even more lysate could be used to web or calculate the titer.