IGEM:The Citadel/Notebook/Notes Brian/05-30-10

May 30

• Prepared 500mL of SOB for the Electroporation on Monday. Needs to be pH tested and converted to SOC.
• Prepared 500mL of LB Lennox (see below).
• Reviewed protocols for training sessions this coming week.
• Designed a genetic circuit for team's review tomorrow during the long wait for culture growth.

This is a continuation of our work from 05-29-10:

Protocol for Electrocompetent Cells

Procedure

1. Autoclaved all flasks and tubes.
   The 250mL tubes were autoclaved with caps off, so to ensure sterility we've stored them on ethanol-soaked paper.
2. Prechilled the rotor at 4°C.
   All tubes and pipets will be chilled tomorrow morning upon arrival.
3. Inoculated 5mL LB Miller medium and placed in incubator for overnight growth at 37°C with 240 RPM.
   Austin Che calls for LB Lennox to lessen salt content. We had only LB Miller at the time of inoculation and have attempted to use that; we'll post an update! For tomorrow's 450mL culture, we've prepared 500mL of LB Lennox.