Koshland Web Site/Methods
FROGGING
FROGGING
  1. Frog is 8x6 prong. keep in EtOH, flame, then dip in sterile dH2O before use.
  2. Dip & swirl into yeast diluted in microtiter plates.
  3. Place on dry plate 1-2sec, remove, will leave 1-2 ul.
  4. Serial dilutions (in steril water on YPD) -8 samples x 6 dilutions. Aim for initial dilution at ~OD 0.1 [850 cells/uL] in 250 ul. 5X dilutions (do 4) 50 ul into 200 ul H2O
  5. Check plates, sometimes not even. Try pressing on FROG

    OD600 [cells/mL] cells/1 ul spot 5X dilution
    ~0.1 85x104 850 850
    28x104 283 170
    9.4 94 34
    3.2 32 6.8
    1 11
    0.35 3.5
Comments:
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