Overnighting Bacteria

— Jacob Anderson 2018/11/16 13:32

The purpose of this experiment is to propagate a bacterial strain for further experimentation. You may need to do a transformation or run a midi or mini - prep for measure the concentration of DNA. Either way follow these basic steps.

• LB + Drug required for selection
• 37C shaker (usually on the first follow of the CRF near the autoclave)
• Culture tubes
• Flasks twice the size of your expected mixture
• LB OP-50 stock for control

Controls

1. Add 3 mL of LB to each culture tube
2. Add 1.5 mL of Bacteria to each culture tube
3. Add the drug you are using in your experiment to one of the culture tubes
4. PLace in 37C shaking incubator found on the first floor of the CRF near the autoclave.
5. Label tubes with Name of bacteria and media, date, initials, MM-LAB.

Flasks

1. Add 125 mL LB to each 250mL flask.
2. Add calculated amount of Drug to the LB in the flasks.
3. Pull bacterial strain from the -80C freezer and add to each of the flasks.
4. Place flasks in the 37C shaking incubator found on the first floor of the CRF near the autoclave.
5. Label tubes with Name of bacteria and media, date, initials, MM-LAB.