User:Karmella Haynes/Notebook/miR Trigger/2011/06/28

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06/28/11

 * &#x2713; Transfection: miR sensor test set #1 (no Gal4 binding sites), U2OS plain

Transfections: miR sensor set 1, 0 Gal4 BS Plate 1: Lipo LTX > Lipofectamine LTX > Follow Jason's protocol, 12-well plates

--> 37-39. no transfection

Lipo LTX Note: set up 3x vol reactions > Add 180 μL Opti-MEM to 5 μL DNA; 3x = 540 μL Opti-MEM, 15 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


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