******Abbreviated protocol******
Surface area of 10 cm dish is 8 x greater than that of a 6-well dish.
Therefore:
1. Use 20 ug of plasmid DNA.
2. Use 64 ul of Lipofectamine 2000 per dish
3. Place each into 1.2 ml of Opti-MEM.
4. Let sit at RT for 5 minutes.
5. Mix and let stand 20 minutes prior to addition to cells.
6. Cells should be in ~8 ml of media (w/o drugs of any kind).
Continue as usual.
******Abbreviated protocol******
Surface area of 15 cm dish is 18 x greater than that of a 6-well dish.
Therefore:
1. Use 45 ug of plasmid DNA.
2. Use 144 ul of Lipofectamine 2000.
3. Place each into 2.7 ml of Opti-MEM.
4. Let sit at RT for 5 minutes.
5. Mix and let stand 20 minutes prior to addition to cells.
6. Cells should be in ~20 ml of media (w/o drugs of any kind).
Continue as usual.