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Salient

Salient is an excellent design with a fresh approach for the ever-changing Web. Integrated with Gantry 5, it is infinitely customizable, incredibly powerful, and remarkably simple.

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Yeast Media

Plate pouring

To make plates, it is the easiest to prepare 2 L batches. Make one 2L flask with 1L of water and 40g of agar. Take a 1L bottle and use that for the actual medium (so use 2 times the amount for 1L). Make it up in 900 ml of water, to leave room for 100 ml of 40% glucose. Put a stirbar in the bottle. Autoclave the flask and the bottle, and pour the contents of the bottle into the flask. Then add glucose, amino acids etc.

YEPD

For 1L medium:

  • 20g yeast peptone
  • 10g yeast extract
    ----autoclave----
  • add 50 ml 40% glucose

! For ade2 strains such as Y8800 and Y8930, extra adenine needs to be added (150mg/L).

Synthetic Complete (SC) dropout medium

For MaV103/MaV203, use -Leu -Trp -His -Ura amino acid mixture
For Y8800/Y8930, use -Leu -Trp -His -Ade amino acid mixture
Or use -Leu -Trp -His -Ura -Ade, which is suitable for both

For 1L medium:

    • 1.3g AA powder
    • 1.7g YNB (without AA and Ammonium Sulfate)
    • 5g Ammonium Sulfate
    • 150 mg Adenine if needed
    • Adjust pH to 5.9
      ----autoclave----

 

  • add 50 ml 40% glucose
  • add 8 ml of each additional AA you want (Leu, Trp, His, Ura) and 3AT or 5-FOA

SC additives

100 mM Histidine (don't autoclave) (Sigma H8000)
100 mM Leucine (Sigma L8000)
40 mM Tryptophan (keep in aluminum foil at 4°C, don't autoclave) (Sigma T0254)
40 mM Adenine (if Adenine comes out of solution, briefly heat up) (Sigma A3159). Easier is to add the Adenine as a powder (76 mg/L for MaV strains, 150 mg/L for Ade- strains such as the Y strains).
20 mM Uracil (Sigma U0750)

3AT concentrations range from 20 - 100 mM for MaV103/MaV203, and 0-5 mM for Y8800/Y8930

20 mM 3AT = 1.68 g/L
5-FOA = 1g/L or 2g/L
Cycloheximide = 1µg/ml final from a stock of 10mg/ml in ethanol
Canavinine = 60µg/ml from a 60mg/ml stock in water.

The color codes for on the plates

color coding

 

Amino Acid powder

Mix 3g each of the following amino acids. Note that the Sigma numbers in front of each amino acid are only a suggestion. Sigma sells lots of variants of each amino acid, and I don't understand the significance of many of the differences. Most likely, any L-Amino Acid should work fine, and you can get the cheapest. The mixture will remain stable for at least 6 months. Adenine and Uracil are optional, depending on the yeast strain used. If using ade2 yeast strains such as Y8800 or Y8930, use double the amount of Adenine.

Amino acid powder components
Code Name Code Name
A3159
A7627
A5006
A9256
A8381
C7352
G5889
G3126
G7126
adenine sulfate
alanine
arginine
aspartic acid
asparagine
cysteine
glutamic acid
glutamine
glycine
I2752
L5626
M9625
P2162
P0380
S4500
T8625
T3754
U0750
V0500
isoleucine
lysine
methionine
phenylalanine
proline
serine
threonine
tyrosine
uracil
valine