User:Karmella Haynes/Notebook/miR Trigger/2011/05/30

{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

05/30/11

 * &#x2713; Transfection: miR sensor constructs into U2OS plain

Lipo LTX transfection > Follow Jason's protocol, 12-well plates > Set up each in triplicate, 48 samples (4 plates) total > Keep 5th plate as "blank" control for FACS


 * Poor quality DNA (see BioBrick Cloning entry 5/30/11)

> Dilute 1 ug DNA in 20 μL (total) st dH2O; set up 3x and aliquot 20 μL to other 2 tubes > Add 180 μL Opti-MEM > Add 1 μL PLUS reagent --> R.T/ 5 min. > Add 3 μL Lipo LTX --> R.T/ 30 min. > Add ~200 μL complexes (drop-wise) to each well (1 ml med. each); Grow cells at 37&deg;C


 * }