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.
Results
Growth test results of synthetic medium with relevant genotypes (Tables 3 and 4).
References
Pour continuer à lire, veuillez vous connecter à votre compte ou en créer un.
Vous n'avez pas de compte ?