Recipe for low concentration Benomyl plates (0, 2.5, 5, 7.5, 10, 12.5 and 15 ug/ml)

1. Make a 10mg/ml stock of benomyl in DMSO. do not make a higher concentration since it comes out of solution upon addition to YEPD
   
   For 10 plates at each benomyl concentration:
2. place a stir bar into a 500ml flask or bottle
   
3. add 250ml YEPD + 5g Agar
   
4. autoclave and cool to 55^oC in a water bath
   
5. make a series of concentrations of benomyl as follows:
   Place flask onto stir plate. begin stirring and add the appropriate amount of DMSO then add benomyl dropwise
   a) 0ug/ml benomyl (control): add 500ul DMSO (0.2% final)
   b) 2.5ug/ml benomyl: add 62.5ul benomyl stock + 437.5ul DMSO
   c) 5ug/ml benomyl: add 125ul benomyl stock + 375ul DMSO
   d) 7.5 ug/ml benomyl: add 187.5ul benomyl stock + 312.5ul DMSO
   e) 10ug/ml benomyl: add 250ul benomyl stock + 250ul DMSO
   f) 12.5ug/ml benomyl: add 312.5ul benomyl stock + 187.5ul DMSO
   g) 15ug/ml benomyl: add 375ul benomyl stock + 125ul DMSO
   
6. pour plates and allow to dry for 2 days on bench top then store at 4^oC. Don't leave plates on the bench too long. I tested plates left on the bench 4 days vs those on the bench 2 days and found that the 4 day plates were less inhibitory for growth.
   NOTE: I usually make 2X as many control plates because these tend to get used up quicker or get contaminated more readily than the benomyl containing plates.
   
   

Recipe for high concentration Benomyl plates (70ug/ml)

1. Make a 10mg/ml stock of benomyl in DMSO. Do not make a higher concentration since it comes out of solution upon addition to YEPD
   
   Recipe for one liter of plates at 70ug/ml:
2. Place a stir bar into a 2 liter flask
3. Add 1000ml YEPD + 20g Agar
   
4. Autoclave and cool to 60^oC in a water bath
   
5. Place flask onto stir plate on low heat setting. begin stirring then using a P1000 pipetman, add dropwise, 7ml of the 10mg/ml benomyl stock to achieve a final concentration of 70ug/ml benomyl
   
6. Pour plates and allow to dry for 2 days on bench top then store at 4^oC. Don't leave plates on the bench too long. I tested plates left on the bench 4 days vs those on the bench 2 days and found that the 4 day plates were less inhibitory for growth