IGEM:VGEM/2007/Notebook/2007-8-7


 * Transforming DB3.1 Cells:
 * 1) Thaw competent cells on ice.  Place required number of 17 × 100 mm polypropylene tubes on wet ice.
 * 2) Gently mix cells, then aliquot 100 µl competent cells into chilled polypropylene tubes.
 * 3) Refreeze any unused cells in the dry ice/ethanol bath for 5 minutes before returning them to the -80°C freezer.  Do not use liquid nitrogen.
 * 4) Add 5uL of biobrick construction plasmid DNA to cells, moving pipet through cells to mix.  Gently tap to mix.
 * 5) Incubate cells on ice for 30 minutes.
 * 6) Heat-shock cells 45 seconds in a 42°C water bath; do not shake.
 * 7) Place on ice for 2 minutes.
 * 8) Add 0.9 ml of room temperature S.O.C. Medium (Cat. No. 15544-034).
 * 9) Shake at 225 rpm (37°C) for 1 hour.
 * 10) Plate transformed cells @ 75uL and 150 uL on ampicillin plates
 * 11) Plate control cells @ 100uL on control and ampicillin plates (for a total of 8 plates)
 * 12) Incubate overnight at 37°C.


 * Butanol Tolerance
 * Plated 1.6% butanol cells.
 * Added yesterday's 1.6% butanol tolerant cells to 3.2% butanol broth solution.
 * Put 3.2% solution cells in shaker at 37oC overnight