06/02/11
- ✓ 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
- Specimen_001: blank (U2OS plain)
- Specimen_002: blank "
- Specimen_003: KAH183/176
- Specimen_004: KAH183/176 +dox
- Specimen_005: KAH184/176
- Specimen_006: KAH184/176 +dox
- Specimen_007: KAH183/177
- Specimen_008: KAH183/177 +dox
- Specimen_009: KAH184/177
- Specimen_010: KAH184/177 +dox
- Specimen_011: JDS33-47 (no transf.)
- Specimen_012: JDS33-47 (no transf.) +dox
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