User:Karmella Haynes/Notebook/miR Trigger/2011/07/07

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07/07/11

 * &#x2713; Daniel - Transfections: Lipo LTX miR-34 sensors in U2OS plain

Transfections: miR-34 sensors > Lipofectamine LTX > Follow Jason's protocol, 12-well plates

Lipo LTX Note: set up 3x vol reactions > Dilute 1 μg DNA in dH2O (10 μL final); 3x = 3 μg DNA in dH2O (30 μL final) > Add 190 μL Opti-MEM to 10 μL DNA; 3x = 570 μL Opti-MEM, 30 μL DNA > Add 1 μL PLUS reagent --> R.T/ 5 min.; 3x = 3 μL PLUS > Add 3 μL Lipo LTX --> R.T/ 30 min.; 3x = 9 μL Lipo LTX > Add ~200 μL complexes (drop-wise) to each well (1 ml med. each); Grow cells at 37&deg;C

> Microscopy and flow cytometry tomorrow (7/08/11)


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