User:Karmella Haynes/Notebook/Polycomb project/2010/06/24

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06/24/10

  • ✓ Cell culture: expand Pc-ATF lines 129-4(1/23/10), 131-9 (10 cm plates)
  • ✓ Senescence assay: KAH126-1, 126-3, 132-8, 154-2 -/+ dox and FTRx +rotenone
  • ✓ Transformation (bact.): quick and dirty transformation for KAH96, 142, 146, 147 into C2925 (dam-/dcm-); morning
  • ✓ Cultures: pick one colony each, grow 5 mL culture o/n
  • ✓ Transfection prep: seed FTRx plate for H3me sensor stables (try DNA -met plasmids); ~200k cells/ well, 6-well plate



Senescence assay
> Samples (plate ~200k cells/ well, 6-well plates, ~3.5 mL medium/ well):

  1. KAH126-1
  2. KAH126-1 +1 μg/mL dox
  3. KAH126-3
  4. KAH126-3 +1 μg/mL dox
  5. KAH132-8
  6. KAH132-8 +1 μg/mL dox
  7. KAH154-2
  8. KAH154-2 +1 μg/mL dox
  9. FTRx + 0 rotenone (7.0 μL DMSO)
  10. FTRx + 0.1 μg/mL rotenone (1.75 μL/ 5.25 μL DMSO)
  11. FTRx + 0.2 μg/mL rotenone (3.5 μL/ 3.5 μL DMSO)
  12. FTRx + 0.4 μg/mL rotenone (7.0 μL/ 0 μL DMSO)



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