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western blot yeast

  1. spin down the over night culture 2 min 10000 rpm
  2. pour off the SN
  3. add 2x Lämmli: 50 – 100 µl → vortex
  4. boil for 5 min → chill on ice
  5. spin down 30 s top speed→ chill on ice
  6. 15 µl on gel
  7. 2 ml over night culture (selective medium) @ 30°C

pGBKT7: α-c-myc (1:5000)
pGADT7: α-HA (1:5000)
pJG4-5/pJCL: α-HA (1:5000)
pEG202: α-LexA