IGEM:VGEM/2007/Notebook/2007-8-13

Things to accomplish

 * 1) Order top system/ cellulase system
 * 2) Discuss GCMS standards
 * 3) Design fermentation experiment

Things accomplished

 * 1) Evaluate remaining funds
 * 2) Continue butanol experiment
 * 3) Placed colony from 3.2% butanol plate into 2.4% butanol broth
 * 4) Maxi prep construction vectors (A/K, A/C)
 * 5) Re-transform failed vector (A/T)
 * 6) * Transforming DB3.1 Cells:
 * 7) Thaw competent cells on ice.  Place required number of 17 × 100 mm polypropylene tubes on wet ice.
 * 8) Gently mix cells, then aliquot 100 µl competent cells into chilled polypropylene tubes.
 * 9) 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.
 * 10) Add 5uL of biobrick construction plasmid DNA to cells, moving pipet through cells to mix.  Gently tap to mix.
 * 11) Incubate cells on ice for 30 minutes.
 * 12) Heat-shock cells 45 seconds in a 42°C water bath; do not shake.
 * 13) Place on ice for 2 minutes.
 * 14) Add 0.9 ml of room temperature S.O.C. Medium (Cat. No. 15544-034).
 * 15) Shake at 225 rpm (37°C) for 1 hour.
 * 16) Plate transformed cells @ 75uL and 150 uL on ampicillin plates
 * 17) Plate control cells @ 100uL on control and ampicillin plates (for a total of 8 plates)
 * 18) Incubate overnight at 37°C.
 * 19) Finish E. coli model