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  • Common Yeast Media (YPD, SC, and the like)
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YPD liquid media (and plates)

  1. Mix in < 1L water:

    Bacto-yeast extract10g
    Bacto-peptone20g
    (Bacto-agar if making plates)(20g)
  2. Bring to 950ml.

  3. Autoclave 15 min at 121C.
  4. Let it start to cool, and add 50ml of 40% sterile glucose solution. Swirl to mix.
  5. (Pour plates.)

YPGal liquid media (and plates)

  1. Mix in < 1L water:

    Bacto-yeast extract10g
    Bacto-peptone20g
    (Bacto-agar if making plates)(20g)
  2. Bring to 950ml.

  3. Autoclave 15 min at 121C.
  4. Let it start to cool, and add 50ml of 40% sterile galactose solution. Swirl to mix.
  5. (Pour plates.)

YPRaf liquid media (and plates)

  1. Mix in < 1L water:

    Bacto-yeast extract10g
    Bacto-peptone20g
    (Bacto-agar if making plates)(20g)
  2. Bring to 950ml.

  3. Autoclave 15 min at 121C.
  4. Let it start to cool, and add 50ml of 40% sterile raffinose solution. Swirl to mix.
  5. (Pour plates.)

SC (synthetic complete)

From CSH methods, for 1L total:

  1. Mix into 900 or 950ml H20 (see step 3).

    YNB (w/out ammonium
       sulfate or amino acids)
    1.7g
    Drop-out mix (e.g. SC-ura-trp)2g
    Ammonium Sulfate5g
    Bacto agar (to make plates)20g
  2. Autoclave, let cool.
  3. Add 20g glucose, say 50ml of 40% glucose or 100ml of 20% glucose.

Using G418 plates? Ammonium sulfate inhibits this selection, so you're going to have to substitute in 1g/L of L-glutamate. However, glutamic acid changes the pH to degrade the agar, so you'll also have to bring the pH back up using a bit of NaOH. ~ 1 of the smaller pellets seems to work, a stack of plates was pH 6.5 to 7 by test strips and was poured as beautiful solid plates.

SC+(5-FOA) Plates (1 Liter, ~2 sleeves)

Method 1:
  1. Add 20g agar to 500ml diH20 in  large flask (>1L, see step 4), autoclave for 15-20min sterilize time.

  2. Combine:

    YNB w/o ammonium sulfate w/o aa1.7g
    Ammonium sulfate5g
    Drop-out mix (-ura)2g
    Uracil (of 2g/L pre-mix)25ml
    5-FOA (0.1% final conc)1g

    Bring to 500ml with diH20. Filter sterilize, 0.2 micron filter.

  3. Let agar cool to ~80C.

  4. Add filtered solution to agar solution, mix carefully.
  5. Pour plates.
Method 2:
  1. Dissolve 1g 5-FOA in 10ml DMSO (this takes a hour or two). Also requires dissolved glucose solution, 20% used below, or adjust for using 40% glucose.
  2. Dissolve into 865ml diH20:

    YNB w/o ammonium sulfate w/o aa1.7g
    Ammonium sulfate5g
    Drop-out mix (-ura)2g
    Bacto-agar20g

    Autoclave liquids setting, 15min sterilize.

  3. Let autoclaved SC base media cool until you can hold the base of the flask in your hand without severe pain. ~65C.
  4. Add 100ml of 20% glucose for 2% final conc.
  5. Add 10ml of the 100g/L 5-FOA solution, 1g total 5-FOA for 0.1% final conc.
  6. Add 25ml of 2g/L uracil solution in H20 for 50ug/ml final conc.
  7. Swirl, let settle, pour plates.
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