Skatebro:Pouring Plates

1. Set water bath to 50 degrees
2. Preparation of LB Agar
   • Obtain 1.2% LB agar from the media room (500 ml per bottle).
     • Be sure to sign it out
   • Melt in microwave:
     • loosen the cap a lot, lay down paper towels in case of spill, use 50% power (enter time, press Power, 5, Start)
     • monitor as you melt, takes approx. 10 minutes per bottle (if doing 2 or 3 bottles simultaneously, add 5 minutes)
     • If too hot, let the melted solution cool so that it's warm, but not hot to the touch - you should use the water bath to do this (50-60 degrees) so it won't resolidify.
   • Once it's cooled to 50-60 degrees then add antibiotics (and label LB jars accordingly):
     • For 500mL of LB Agar use:
       • 500μL Amp stock (Amp stock = 50mg/ml)
       • 1mL Kan stock (Kan stock = 10mg/ml)
       • 750μL Gent stock
       • 500μL Tet stock
       • 500μL Chl stock
     • Swirl to mix; try not to make too many bubbles.
   • Pour
   • Leave plates to solidify on benchtop (overnight even).
   • Label each plate to indicate antibiotic.
   • Store the plates in their original bags in fridge