Cultivation and Transformation of Yeasts. Tian He 2008. 07. 06. Media. non-selective: YPD without antibiotics Selective: Synthetic complete dropout medium (SC) (auxotroph) YPD with antibiotics. YPD: 1000 mL Yeast Extract 10 g Peptone (Tryptone) 20 g
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2008. 07. 06
YPD without antibiotics
Synthetic complete dropout medium (SC)
YPD with antibiotics
Lower: yeasts grow slowly.
Higher: yeasts die.
LEU2, URA3, TRP1, HIS3, ADE1,2
pGREG 503 504 505 506
HIS TRP LEU ADE
Question: why we use pGREG503 for homologous recombination?We cannot determine from the phenotype whether recombination occurs.
Autonomously replicating sequence
GAL 4 induced promoters: GAL1, GAL2, MEL1
The competent cells of yeast cannot be stored;
it must be prepared before use!
centrifuge for 30 s at 13.000 g and discard the supernatant.