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

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06/06/10

  • ✓ ChIP: order primers for ChIP-DNA PCR
  • ✓ Senescence assay optimization: FTRx cell rotenone treatment
  • ✓ Growth assay: 12-well plates for growth assay (from 10cm plate cultures -/+ dox)


ChIP Primers
> Genes (see spreadsheet for sequences)

  1. p15 INK4b
  2. p14 ARF
  3. p16 INK4a
  4. ATF3
  5. CCND2
  6. GAPDH


Senescence assay
> FTRx cell rotenone treatment
> 12-well plate; ~100k cells/ well

  • 1-4: 0, 0.1, 0.2, 0.4 μg/mL rotenone
  • 5-8: 0, 0.1, 0.2, 0.4 μg/mL rotenone
  • 9-12: 0, 0.1, 0.2, 0.4 μg/mL rotenone

Note: Each row will be treated w/ different concentrations of C12-FDG
> Grow at 37°C/ 3 days
> Next step: Wed, 6/09; C12-FDG treatment & microscopy


Growth assay
> 12-well plates; ~100k cells/ well
Plate 1, 2

  • 1-4: 126-1
  • 5-8: 126-3
  • 9-12: 132-8
  • 13-16: 154-2
  • 17-20: FTRx

Plate 3, 4

  • 1-4: 126-1 +dox
  • 5-8: 126-3 +dox
  • 9-12: 132-8 +dox
  • 13-16: 154-2 +dox
  • 17-20: FTRx +dox

> Refresh dox+ today and every 2 days
> Do cell counts tomorrow and every 2 days


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