Preparing S. Pombe media

Glucose degrades rapidly in the autoclave and will cause variations in cell growth rates. Its best to prepare YES media as follows:

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 0.2µM 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.