I have been having a strange issue growing AH109 and using it for transformation. Usually, I would plate it in YPDA and culture it at YPDA too, the generic protocol that is used for a two-hybrid transformation. However, as I plate it in -LT plates, I get a vigorous growth of pink coloured yeast. I have been unable to get the white colonies at all for some time and I have changed bottles of media, amino acid dropouts and even laminar airflows. I am attaching pictures of the same. I even tried transferring them in -alth x@gal plates but there too I got pink growth. Please help!
Looks like your adenine isn't good anymore (or there was a strain mixup) , AH109 is normally a creamy white color in the presence of adenine. Any chance this wasn't the right strain? Does your lab grow other sorts of yeast?
Maybe borrow/make some -LT plates from a new source?
I used to add in the extra amino acids after autoclaving, maybe that would help?
Maybe borrow some AH109 from another lab? Those colonies look just a bit odd.
Amy Klocko I got the AH109 strain from another lab and this issue is still persisting. Also got-LT plates from someone else, but at that time no cells grew at all. Even added extra adenine after autoclaving. I will try to get some adenine from another lab. Hopefully, it will solve this issue.
I'd try another source for the AH109 yeast. That looks like the Ade-mutant phenotype.
But I got this from the TakaraBio user manual:
"In the absence of GAL4, AH109 also exhibits the ade2-101 phenotype. However, in the presence of protein interactions, the ADE2 marker complements in cis the AH109 ade2-101 phenotype."
Yeast strains are pretty cheap. I would just buy a fresh stock from a supply company. You don't want to spend weeks or months on this project only to start over due to using the wrong yeast strain.
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