Team:Brown/Notebook Recipes/TSBPlates

From 2009.igem.org




Making Tyrpticase Soy Broth (TSB) Plates (For 1L = ~ 30plates)

1. Add 30g of TSB into 2. 1 L ddH2O in 2L of Flask 3 Swirl the flask until all the LB is dissolved. 4. Add 15g Agar 5 Bring the solution to a boil (Clear) on a hotplate with a stir bar stirring. (Temperature ~ 430 oC) 6 Autoclave the solution at 20min at 15psi and 121 oC. 7 Cool the temperature down to 50 oC - 60 oC OR cool to the touch. 8. 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] 9. Swirl the solution slowly to mix the antibiotics without forming a lot of bubbles. 10. 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.

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