Electrocompetent yeast

The final volume of resuspended, electrocompetent yeast should be 1-1.5 mL (rrrNote 5). 

In a sterile, ice-cold Eppendorf tube, mix 40 pL concentrated, electrocompetent yeast with 10-100 ng of transforming DNA (<5 pL) (see Note 6). 

Electrocompetent yeast can be stored at -80°C for future use by adding glycerol to 15% final concentration and snapfreezing in a dry ice/ethanol bath. To use frozen cells, thaw slowly on ice, pellet the cells, and resuspended to the same volume in fresh, ice-cold 1M sorbitol. This will remove excess ions released by dead cells. The transformation efficiency using these frozen/thawed cells will drop > 10-fold from fresh yeast. 

Mixing the DNA with the electrocompetent yeast and allowing them to incubate together on ice for 30-45 min may improve transformation efficiency. 

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See also in sourсe #XX -- [ , , ]

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