Team:Brown/Notebook Recipes/LBPlate


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Making LB Plates (For 1L = ~ 30plates)

1. Add 20g LB into 2. 1 L ddH2O in 2L of Flask o Swirl the flask until all the LB is dissolved. 3. Add 15g Agar o Bring the solution to a boil (Clear) on a hotplate with a stir bar stirring. (Temperature ~ 430 oC) o Autoclave the solution at 20min at 15psi and 121 oC. o Cool the temperature down to 50 oC - 60 oC OR cool to the touch. 4. Add the selected antibiotic (the right concentration and the right volume) o Ampicillin (Amp) = 1 ml of [50ug/ml] o Kanamycin (Kan) = 1 ml of [50mg/ml] o Tetracycline (Tet) = 1 ml of [12.50mg/ml] o Chloramphenicol (Ch) = 1ml of [10mg/ml] 5. Swirl the solution slowly to mix the antibiotics without forming a lot of bubbles. 6. Pour the medium into the dishes as fast as you can without forming bubbles. o Stack the plates on top of each other (~ 5 plates a stack) to lessen the chance of wetting the plates due to condensation. o Flame the mouth of the flask to prevent contamination.