User:Karmella Haynes/Notebook/miR Trigger/2009/12/15

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12/15/09

 * &#x2713; Transfection: RFP reporters into U2OS JDS37-43 (luc miRNA-YFP)
 * &#x2713; Split cells: U2OS plain (1x 1:10)

Transfections > U2OS JDS37-43 (luc miRNA-YFP) cells + RFP-luc target reporters > Use Lipofectamine, 6-well format

> Add Lipo to 250 μL Opti-MEM --> R.T/ 5 min. > Add DNA to 250 μL Opti-MEM > Add DNA mix to Lipo mix --> R.T./ 20 min. > Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37&deg;C > After 5 hours, change medium (ab-free) and add 1 μg/mL doxycycline to "Dox" samples > Check RFP & YFP expression (microscopy) after 18-24 hours


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