How to make LBS Agar medium
Medium: LBS Agar Approximate Formula* 500mL
Ingredients:
Pancreatic Digest of Casein ....................................... 5.0 g Yeast Extract ............................................................... 2.5 g Monopotassium Phosphate......................................... 3.0 g Ammonium Citrate ..................................................... 1.0 g Dextrose ................................................................... 10.0 g Polysorbate 80 ............................................................20 drops Sodium Acetate Hydrate ........................................... 12.5 g Magnesium Sulfate ................................................. .2875 g Manganese Sulfate ..................................................... 0.06 g Ferrous Sulfate .......................................................... .017 g Agar ......................................................................... 7.5 g Water .........................................................................500mL Glacial acetic acid .................................................... 0.66 mL
Materials
- Stir bar
- Stir plate
- Plates
- Clear plastic containers
- Scale
- 1-Liter Jar
To make an LBS medium, using a scale, weigh out the dry ingredients from pancreatic digest of casein to agar using a new clear plastic container for each powder and add it in a 1-liter jar. After adding the dry ingredients in the jar, add 500mL of purified water with a stir bar and place the jar on a stir plate to mix. Now, wait until the dry ingredients have dissolved in the water; about 15 mins. After dissolving the dry ingredients in the water, add .66 mL of glacial acetic acid (ACS) into the jar and then place the jar back on the stir plate at 200 degrees to stir. Keep the jar on the heated stir plate until the temperature of the liquid reaches 100 degrees; about 30 mins. After reaching 100 degrees, allow the liquid to cool for 1 min before placing it into the autoclave with a bottle cap; not too tight. After autoclaving the jar for an hour and a half, allow the liquid to cool for 5 mins before pouring plates.
