Helene Nguewou/Overnight culture

Materials

• Plate of cells to be made competent
• LB media

Glassware & Equipment

• Falcon tubes (cell culture tubes)
• 1ml pipette

Procedure

• Label the tube with your initials, content and date.
• Prepare 5ml of LB solution with the specific antibacterial (Note, CSHL 140 and 141 have no antibacterial resistance, and I 6030 has ampicillin resistnace).
• Use a sterile pipet to scrape a single colony of the agar plate.
• Distribute the colony in the 5ml LB solution by moving the tip of the pipet.
• Put some parafilm on the agar plate and store it at 4 °C.
• Put the media containing the bacteria overnight in the incubator.

• Ideally the cells should not grow for more than 16 hours.

• If a cell with an antibacterial resistance is grown in a media with no antibacterial resistance, the cell will divide and one daughter cell might have the antibacterial resistance and the other might not. With time the cells will loose their antibacterial resistance because the enzyme created by it is not used. Process of adaptation.