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Merck

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.

ComponentAmount per litre of medium (mg)*
1Adenine
18
2p-Aminobenzoic acid8
3Leucine380
4Alanine76
5Arginine
76
6Asparagine76
7Aspartic acid76
8Cysteine
76
9Glutamic acid
76
10Glutamine76
11Glycine76
12Histidine76
13myo-Inositol76
14Isoleucine76
15Lysine76
16Methionine76
17Phenylalanine76
18Proline76
19Serine76
20Threonine76
21Tryptophan76
22Tyrosine76
23Uracil76
24Valine76
Table 1.
* Excluding dropped-out components
Yeast Synthetic Drop-out Medium SupplementAmount per Liter of Medium
Y1501: without uracil1.92 g
Y1751: without histidine1.92 g
Y1896: without lysine1.92 g
Y1376: without leucine1.62 g
Y1876: without tryptophan1.92 g
Y0750: without leucine and tryptophan1.54 g
Y1771: without uracil, leucine, and tryptophan1.46 g
Y2146: without histidine, leucine, and tryptophan1.46 g
Y2021: without histidine, leucine, tryptophan, and adenine1.39 g
Y2001: without histidine, leucine, tryptophan, and uracil1.39 g
Table 2.

Results

Growth test results of synthetic medium with relevant genotypes (Tables 3 and 4).

Synthetic mediumY1501 without uracilY1751 without histidineY1896 without lysineY1376 without leucineY1876 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++-++
Table 3.+ = growth; - = no growth
Synthetic mediumY0750: without Leu & TrpY1771: without Ura, Leu, & TrpY 2146: without His, Leu, & TrpY2021: without His, Leu, Trp, & AdeY2001: without His. Leu, Trp, & UraYPD
       
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++++++
Table 4.+ = growth; - = no growth
Materials
Product No.Product NameDescriptionPricing
Y1896Yeast Synthetic Drop-out Medium Supplementswithout lysine

References

1.
Kaiser C, Michaelis, S., Mitchel, A. 1994. Methods in Yeast Genetics, a Cold Spring Harbor Laboratory Manual , (Cold Spring Harbor, NY).
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