User:Karmella Haynes/Notebook/Polycomb project/2010/11/01

{| 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"|

11/01/10

 * &#x2713; HEKGal4EED + PcATF's: split cells and do luc assay
 * &#x2713; ChIP: H3K27me3 IP for expt. samples (overnight)
 * &#x2713; Transfection: HepG2's (Pc-ATF set + pcDNA6/TetR)
 * &#x2713; Frozen stocks: transfer HepG2's and HEK293's to liq. nitrogen tank.

Lipo transfection > Co-transfect Pc-ATF with pcDNA6/TetR tet repressor plasmid (Invitrogen)

> Add Lipo to 250 μL Opti-MEM --> R.T/ 5 min. > Add Pc-ATF and pcDNA6 plasmid DNA to 250 μL Opti-MEM > Add DNA mix to Lipo mix --> R.T./ 20 min. > Add 500 μL complexes to each 3.5 mL well (4 ml med. each); Grow cells at 37&deg;C


 * }