17.5 grams of LB Agar powder makes 500 mL.
1. Weigh 17.5 grams of LB Agar powder, add into sterilized Pyrex container.
2. Fill with sterile water up to 500 mL mark.
3. Cap loosely and tape with autoclave tape.
4. Autoclave for 40 minutes (see an instructor).
5. Wait until cool enough that it doesn't hurt to touch.
If starting from solid agar in the fridge, heat in microwave (cap unscrewed) 3min on high, 5 min on 30% (till it melts completely)
Wait till cool enough so that you can touch it for a minute (60 degree Celsius)
6. Add appropriate volume of Amp or Kan (1000X). For 500 mL of LB Agar, add 500 uL of each antibiotic needed.
7. Swirl to mix, use pipette to transfer 20 mL into each plate. Avoid bubbles.
8. Allow plates to cool and solidify. Mark antibiotic on side of plate.
If LB Agar solidifies before being poured, microwave to liquefy again.