User:Karmella Haynes/Notebook/miR Trigger/2011/01/23

{| 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"|

01/23/11

 * &#x2713; Transfection: test RFP-repressors
 * &#x2713; Culture: thaw JDS33-47

Lipo transfection > RFP-Repressors with new BGH polyA (from Mara) cloned in forward and reverse orientations in MV8/zeo vector (cloned by Andrew) > Vector has BGH polyA in fwd orientation; RFP expression from reverse clones will show that BGH polyA BioBrick works

> Change medium to ab-free (3.5 mL) > 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 final vol.); Grow cells at 37&deg;C


 * }