Preparing S. Pombe media

Glucose degrades rapidly in the autoclave and will cause variations in cell growth rates. It is best to prepare YES media as follows (see also Microbial Media Recipes):

Contents

1. YES Broth
2. YES plates

1. YES Broth

Contents for 1L:

• 5 g YE (Yeast Extract)
• 30 g D(+) Glucose
• 225 mg Adenine
• 225 mg L-Leucine
• 225 mg Uracil

Add double distilled water to 1 L, mix well with stir bar and sterilize by filtering with a 0.2 µm vacuum filter cup.

2. YES plates

1. Autoclave 20 minutes at 110°C a 1L glass bottle containing 500 ml double distilled water, 20g of agar and a stir bar.
2. Cool down the autoclaved media at 60°C, add 500 ml of filtered 2x YES Broth and mix well with stir bar.
3. Pour 25 ml per Petri dish. Let dry O/N. Store at 4°C in the dark.