Editing ElectroCompetent Cells (section)

== Preparation of Bacterial Cultures, Tools, and Reagents (DAY 1, Afternoon) ==

<br/>1. In the late afternoon, inoculate 1-5 ml autoclaved'''LB broth''' in sterile (for example, 100 x 13 mm) borosilicate glass test tubes with a small&nbsp;aliquot of bacteria (E. coli or Vibrio natriegens).

2. Set the inoculated test tubes in a roller drum housed at 37 °C temperature (warm room or incubator), turn on the roller drum at high speed, and incubate O/N.

<br/>3. Prepare a cell spreader in the shape of a "hockey stick" from a glass rod by heating the middle of the rod in the flame of a Bunsen burner<br/>until the glass softens and then direct the bend gently with tweezers or pliers into a 135° angle. Heat the rod once more in the middle point<br/>between the end and the first bend and bend it at a 45° angle inwards (in the opposite direction of the first bend).

4. Prepare LB-agar without antibiotics and pour into Petri dishes in preparation of the electrocompetence protocol, and LB-agar plates<br/>containing the appropriate antibiotic (according to the resistance marker on the vector to be electroporated), and store at 4 °C.

5. Prepare a filter sterilized '''2 mM CaCl2''' solution for Vibrio natriegens, or autoclave ddH2O for E. coli, and store at 4 °C.