Team:Aachen/Wetlab/Protocol27

S. cerevisiae minimal medium with synthetic dropout complement:

[URA-]-medium with synthetic drop-out complement:(1000 ml)


  • 6.7 g nitrogen base without AA.
  • 40 ml 50% glucose solution.

  • 1.92 g synthetic drop-out complement (Sigma Alderich).

Procedure:


  1. Dissolve the reagents in 960 ml deionized water (800 ml if preparing medium containing raffinose).
  2. If you are making plates, add the agar after dissolving the reagents above.
  3. Autoclave at 15 psi, 121 ℃ for 20 minutes.
  4. Cool to 50 ℃ and add 100 ml of filter-sterilized 20% glucose or 200 ml of filter-sterilized 10% raffinose.
  5. Pour plates and allow to harden. Invert the plates and store at 4 ℃. Plates are stable for 6 months.

[HIS-]-medium with synthetic drop-out complement:(1000 ml)


  • 6.7 g nitrogen base without AA.
  • 40 ml 50% glucose solution.

  • 1.92 g synthetic drop-out complement (Sigma Alderich).

Procedure:


  1. Dissolve the reagents in 960 ml deionized water (800 ml if preparing medium containing raffinose).
  2. If you are making plates, add the agar after dissolving the reagents above.
  3. Autoclave at 15 psi, 121 ℃ for 20 minutes.
  4. Cool to 50 ℃ and add 100 ml of filter-sterilized 20% glucose or 200 ml of filter-sterilized 10% raffinose.
  5. Pour plates and allow to harden. Invert the plates and store at 4 ℃. Plates are stable for 6 months.

[URA-]-solid-medium with synthetic drop-out complement:(1000 ml)


  • 6.7 g nitrogen base without AA.
  • 40 ml 50% glucose solution.

  • 1.92 g synthetic drop-out complement (Sigma Alderich).

  • 20 g Agarose.


Procedure:


  1. Dissolve the reagents in 960 ml deionized water (800 ml if preparing medium containing raffinose).
  2. If you are making plates, add the agar after dissolving the reagents above.
  3. Autoclave at 15 psi, 121 ℃ for 20 minutes.
  4. Cool to 50 ℃ and add 100 ml of filter-sterilized 20% glucose or 200 ml of filter-sterilized 10% raffinose.
  5. Pour plates and allow to harden. Invert the plates and store at 4 ℃. Plates are stable for 6 months.