5-FOA AGAR (1L)

Mix A (in 2L flask w stir bar)

Bacto-agar 20 gm

Yeast N-Base 6.7 gm

H2O to 500 mL

Mix B

Amino Acid Powder (complete)   1 gm

5-FOA   1 gm

Glucose (40% w/v) 50 ml

H2O 450 ml

Make Mix B and stir on heat plate on low (approx 65 C; use thermometer)

Takes approx. 20-25 min for 5-FOA to go into soln

When powders are fully dissolved add glucose

Filter sterilize

While Mix B is dissolving, autoclave Mix A 25 min

When autoclaving is done add sterile Mix B immediately and stir well

Notes:

1)  To scale up to 2L of plates autoclave agar in 4L flask

2)  Can make plates missing amino acids by adding appropriate drop out powder

3)  Can substitute galactose for glucose if needed.

4)  Mix A is 4% agar. It will have a greater tendency to bump and will solidify faster than 2%.  That's why Mix B is added without letting Mix A cool.

5)  These plates appear to work for up to 9 mos if stored at 4 C (keep in dark)