Hi there,

I don't think there is anything specific to yeast transformation. If you want to compare different transformations they have to be done on the same cells using the same reagents and at the same time. For more info about parameters influencing yeast transformation efficiency, have a look at the following:

Article Transformation of Saccharomyces cerevisiae and other fungi

For reagent preparation please consult the Gietz lab page (even if it is not updated!):

https://home.cc.umanitoba.ca/~gietz/

Hi. Why not to try LiAc? It's efficiency is higher as compared.

Please check this paper:

https://link.springer.com/article/10.2478/s11535-008-0040-7

You have to consider cell-to-cell heterogeneity of metabolic states in isogenetic populations.

"The results of our study showed that the fluctuation in transformation efficiency was strictly dependent on the metabolic state of yeast cells and the capacity of the yeast cells to become competent was related to the S-phase of cell cycle" " The transformation efficiency of yeast cells during exponential growth might be characterised as undulatory. The aim of the study was to investigate the reason for the fluctuation in transformation efficiency of yeast Saccharomyces cerevisiae p63-DC5 cells during exponential growth"

LiAc/PEG transformation is very standard, transformation efficiency is reliant on the cells bring in mid exponential phase and gentle handling. In general it’s the ligation efficiency which is more important, make sure your using a 1:3 vector:insert. As a rule of thumb I’ve always found selection based on antibiotic resistance works better.

I don’t know which yeast your using but I tend to use BY4741 or W303.

There are ways of improving efficiency such as using a PCR based approach to cloning and the relatively new takara vectors.

Hope this is of any use.

Thanks all for useful tips. I am going to try suggestions made here.