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

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06/02/11

 * &#x2713; 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


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