User:Karmella Haynes/Notebook/miR Trigger/2011/06/02

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  • ✓ Flow cytometry: HPK/HPK miR sensor set in JDS33-47

Flow cytometry
> Checked transfected cells for RFP/CFP. CFP is easy to see, RFP very dim/ infrequent
> Added 1μg/mL dox to second well of each sample pair @ 11am
> Harvested cells for flow at 1:30 pm (~2 hours miR/YFP activation)

> Use 96-well round-bottom plate

  • Trypsinize cells w/ 0.5 mL trypsin medium
  • Harvest using 1 mL plain medium, transfer 1.5 mL cells to 15 mL conical
  • Spin at 1100 rpm @ 4°C
  • Pour off sup. and resuspend cells in 200 μL 1x PBS, transfer 200 μL to 96-well plate

> Experiment/folder: Karmella mCherry AmCyan Venus > 060111 MS Repression

  1. Specimen_001: blank (U2OS plain)
  2. Specimen_002: blank "
  3. Specimen_003: KAH183/176
  4. Specimen_004: KAH183/176 +dox
  5. Specimen_005: KAH184/176
  6. Specimen_006: KAH184/176 +dox
  7. Specimen_007: KAH183/177
  8. Specimen_008: KAH183/177 +dox
  9. Specimen_009: KAH184/177
  10. Specimen_010: KAH184/177 +dox
  11. Specimen_011: JDS33-47 (no transf.)
  12. Specimen_012: JDS33-47 (no transf.) +dox