#Begin by obtaining two tubes of LB broth (each will have 5 ml of broth) from the refrigerator in the back left hand corner of the room.<br>
#Add 5 microliters of the 50mg/ml ampicillin stock (also found in the refrigerator) to each tube. '''Calculate the effective concentration of ampicillin''' that you will have in your LB tube (remember V1 x C1= V2 x C2) and record that information in your lab notebook. <br>
#Gently swirl your LB +amp broth to mix the contents. <br>
#Label the two sterile glass culture tubes with tape in your team color. Label one with "pPD129.36 ''lsy-2''" and your initials. Label the other with your initials only. <br>
#Inoculate the broth with your bacteria by using a sterile disposable loop
or sterile toothpick to scrape into the broth the rest of a colony that was positive for evidence of our plasmid by colony pcr. Be sure not to touch the plate with the loop or toothpick except on the desired colony and don’t pick up any satellite colonies! Gently swirl the loop or toothpick in the LB+amp broth - you should be able to see the colony come off into the broth. (The second tube of broth labeled with just your initials is a control and should not be inoculated with bacteria.) <br>#Balance the 2 tubes across from each other on the rotating wheel in the 37C incubator at the front of the room when you come in the door. DO NOT USE THE ROTATING WHEEL AT ROOM TEMPERATURE!
#Incubate these broth cultures at 37°C overnight. '''Do not forget to make sure the wheel is rotating when you leave!'''<br>