User:Karmella Haynes/Notebook/Polycomb project/2009/12/01

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12/01/09

 * Transfections: mCh-VP64 (trial 3)

Transfections > HEK293 cells + Gal4/mCh/VP64 constructs > Use Fugene, 24-well format > Serial dilutions of plasmids --> Start with 1600 ng in 10 μL OptiMEM --> Add 5 μL plasmid to 5 μL OptiMEM for each serial dilution

> Master mix: 90 μL Fugene + 910 μL Opti-MEM --> R.T/ 5 min. > Add 20 μL Fugene mix to DNA --> R.T./ 20 min. > Add 25 μL complexes to each well (1 ml med. each); Grow cells at 37&deg;C > Assay RFP/ Luc after 18-24 hours


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