Yeast Drop Out Bulletin
The selection of plasmids in yeast is based on the use of auxotrophic mutant strains, which cannot grow without a specific medium component (an amino acid, purine or pyrimidine). Transformation with a plasmid containing the mutated gene enables the transformant to grow on a medium lacking the required component. Although yeast can grow on a synthetic medium without any amino acids, better yield and growth rate can be achieved on richer media. Having a richer medium also increases the probability of successful transformations when screening libraries or performing gene knockouts. Composition of the media supplements is shown in Table 1 and is based on published formulations.1
Precautions and Disclaimer
This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.
Preparation Instructions
Yeast Synthetic Drop-out Medium Supplement (Table 2 shows the appropriate amount for each supplement) |
1.39–1.92 g/L | |
Yeast nitrogen base without amino acids (Product No. Y0626) |
6.7 g/L | |
For plates add: Bacteriological agar (Product No. A5306) |
20 g/L |
When preparing plates, autoclave for 15 minutes only; autoclaving longer will cause agar to become soft. Alternatively, the agar can be autoclaved separately from the Synthetic Drop-out Medium Supplement and a yeast nitrogen base.
After autoclaving add:
Glucose, 50% (w/v) solution 40 mL/L
(prepared from Product No. G7021, sterilized by 0.2 mm filtration)
Storage/Stability
Store media supplements desiccated at room temperature.
Component | Amount per litre of medium (mg)* | |
---|---|---|
1 | Adenine | 18 |
2 | p-Aminobenzoic acid | 8 |
3 | Leucine | 380 |
4 | Alanine | 76 |
5 | Arginine | 76 |
6 | Asparagine | 76 |
7 | Aspartic acid | 76 |
8 | Cysteine | 76 |
9 | Glutamic acid | 76 |
10 | Glutamine | 76 |
11 | Glycine | 76 |
12 | Histidine | 76 |
13 | myo-Inositol | 76 |
14 | Isoleucine | 76 |
15 | Lysine | 76 |
16 | Methionine | 76 |
17 | Phenylalanine | 76 |
18 | Proline | 76 |
19 | Serine | 76 |
20 | Threonine | 76 |
21 | Tryptophan | 76 |
22 | Tyrosine | 76 |
23 | Uracil | 76 |
24 | Valine | 76 |
Yeast Synthetic Drop-out Medium Supplement | Amount per Liter of Medium |
---|---|
Y1501: without uracil | 1.92 g |
Y1751: without histidine | 1.92 g |
Y1896: without lysine | 1.92 g |
Y1376: without leucine | 1.62 g |
Y1876: without tryptophan | 1.92 g |
Y0750: without leucine and tryptophan | 1.54 g |
Y1771: without uracil, leucine, and tryptophan | 1.46 g |
Y2146: without histidine, leucine, and tryptophan | 1.46 g |
Y2021: without histidine, leucine, tryptophan, and adenine | 1.39 g |
Y2001: without histidine, leucine, tryptophan, and uracil | 1.39 g |
Results
Growth test results of synthetic medium with relevant genotypes (Tables 3 and 4).
Synthetic medium | Y1501 without uracil | Y1751 without histidine | Y1896 without lysine | Y1376 without leucine | Y1876 without tryptophan |
---|---|---|---|---|---|
Relevant genotype | |||||
ade2, lys2, trp1, his3, ura3, leu2 [pCEN HIS3] | - | + | - | - | - |
ade2, lys2, trp1, his3 , ura3, leu2 [pCEN TRP1] | - | - | - | - | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2] | - | - | - | + | - |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN URA3] | + | - | - | - | - |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2, pCEN TRP1] | - | - | - | + | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2, pCEN TRP 1, pCEN HIS3] | - | + | - | + | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2. pCEN TRP1, pCEN URA3] | + | - | - | + | + |
leu2, trp1, ura3-52 | - | + | + | - | - |
lys1 | + | + | - | + | + |
Synthetic medium | Y0750: without Leu & Trp | Y1771: without Ura, Leu, & Trp | Y 2146: without His, Leu, & Trp | Y2021: without His, Leu, Trp, & Ade | Y2001: without His. Leu, Trp, & Ura | YPD |
---|---|---|---|---|---|---|
Relevant genotype | ||||||
ade2, lys2, trp1, his3, ura3, leu2 [pCEN HIS3] | - | - | - | - | - | + |
ade2, lys2, trp1, his3 , ura3, leu2 [pCEN TRP1] | - | - | - | - | - | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2] | - | - | - | - | - | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN URA3] | - | - | - | - | - | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2, pCEN TRP1] | + | - | - | - | - | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2, pCEN TRP 1, pCEN HIS3] | + | - | + | - | - | + |
ade2, lys2, trp1, his3, ura3, leu2 [pCEN LEU2. pCEN TRP1, pCEN URA3] | + | + | - | - | - | + |
leu2, trp1, ura3-52 | - | - | - | - | - | + |
lys1 | + | + | + | + | + | + |
Product No. | Product Name | Description | Pricing |
---|---|---|---|
Y1896 | Yeast Synthetic Drop-out Medium Supplements | without lysine |
References
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