Agar Bacteriological Plates
- Need a ratio of 25g LB Broth : 15g Agar : 1L dH2O (For 400mL of dH2O, you need 10g LB, 6g Agar)
- Combine in a 500mL glass Duran bottle with a stir bar
- Autoclave in wet cycle for 20 minutes
- Retrieve immediately!
- Stir at 200rpm for 15 minutes without any heating. You want the solution to cool to about 50°C.
- Once cooled enough to keep your bare palm on the bottle without pain, add antibiotic to a final concentration of:
- Kanamycin at 50μg/mL
- Ampicillin 100μg/mL
- Note: Use or make a 1000X stock solution in dH2O and store in the fridge.
- IMPORTANT we use 1000X stock solutions. Therefore in a 400mL bottle of LB:Agar you put in 400μL of antibiotics!!!!
- Pour liquid into sterile 100mm bacterial culture plates (NOT the 100 mm mammalian cell culture plates) to a thickness of about 0.5-1.0 cm, ensuring absolutely NO bubbles remain
- Mark the bottom of the plate with K or A for antibiotic
- Let plates set for a few hours or overnight on bench top (fume hood will dry them out).
- Place them in a bag and store in fridge up to about 3 months. Try to store upside down (bottom facing up) to prevent dehydration.