Superfect Transfections
Dominik Haudenschild September 8, 1999

Culture Cells Media DNA Vol. SF GM
96-well 1e4 100µl 0.5µg 30µl 2.5µl 150µl
48-well 2e4 150µl 0.7µg 50µl 4.5µl 250µl
24-well 4e4 350µl 1.0µg 60µl 5.0µl 350µl
12-well 1e5 800µl 1.5µg 75µl 7.5µl 400µl
6-well 2e5 1.6ml 2.0µg 100µl 10µl 600µl
60mm 4e5 4.0ml 5.0µg 150µl 25µl 1000µl
100mm 2e6 7.0ml 10µg 300µl 50µl 3000µl

1. Seed appropriate number of cells into culture vessel
   
2. Dilute DNA into correct Vol. of media without serum or antibiotics
   
3. Add correct amount of SuperFect SF reagent
   
4. Incubate at RT for 5-10 minutes
   
5. During incubation wash cells with PBS
   
6. Add correct amount of growth media GM to DNA/SF,
   mix by pipetting twice
   immediately add to cells
   
7. Transfect cells for 2-3 hours (1-16 to optimize)
   
8. Wash cells with PBS
   
9. Add growth media, analyze cells after 24-48 hours