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)
- p15 INK4b
- p14 ARF
- p16 INK4a
- ATF3
- CCND2
- 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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