Make LB agar plates for growing bacteria

5 g Tryptone
2.5 g Yeast extract
5 g NaCl

Bring to 500 mls with Millipore water
While mixing, prepare a 1-liter flask with 7.5 g of agar
Adjust LB to pH 7.5 with dilute NaOH
Pour liquid into flask with agar
Cover with aluminum foil and secure with autoclave tape
Autoclave 30 min

Allow to cool with slow mixing to about 50C (should be able to leave your hands on the glass without it hurting)
Meanwhile, remove 1 sleeve of plates from the plastic and save the plastic
Add antibiotic and mix again for a few minutes (Ampicillin at 100 ug/ml, Kanamycin at 50 ug/ml, or Chloramphenicol at 25 ug/ml)
Pour into agar plates, pouring just a little more than it takes to cover the bottom of the plate
Leave at room temperature overnight to reduce condensation
Store inverted in platic sleeves at 4C (refrigerator)

Comments

Popular posts from this blog

Prepare 6X Laemmli buffer

0.1 M Triethanolamine

Laboratory Maintenance Checklist