Abstract
293T and Pheonix cells grow in DMEM + 10%FBS. If you are transfecting other cells, you can use whatever medium those cells normally grow in and change to DMEM + 10%FBS on the day of the transfection. You can change back to “normal” medium 24 hours post-transfection. Calcium Phosphate transfection of M2182 in RPMI (w/o FBS) was reported to cause cells to die. All the transfection have been done in DMEM + 10% FBS, no transfection in other media was performed so far.
Materials and Reagents
Equipment
Procedure
Day 0 Plate 4.5 x 106 293T (or Pheonix for retrovirus) on 10 cm plate. Incubate overnight at 37 °C . You can also scale down to 6 cm or 6 well plates. Note: Alternatively, you can plate the cells the same day as the transfection. See Day 1.
Day 1: Transfection Note: If you plated cells on the previous day, change medium to DMEM + 10% FBS 2 h prior to transfection and skip to step 4.
Recipes
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