The presence of Saccharomyces cerevisiae DNA in various media used to propagate yeasts and its removal by ethidium monoazide.
OBJECTIVE
In this study we demonstrate the interference of yeast extract in enumeration of Saccharomyces cerevisiae using real-time PCR and develop a method for its removal from the media using ethidium monoazide (EMA).


RESULTS
Using real-time PCR and primers to S. cerevisiae we demonstrate the presence of yeast DNA in various media as well as the media impact on S. cerevisiae real-time PCR standard curves. By pretreatment with EMA, we were able to remove this interference.


CONCLUSIONS
Saccharomyces cerevisiae DNA can be found in a number of common laboratory media and may impact the enumeration of this yeast by real-time PCR. However, pretreatment with EMA eliminates this concern.


CONCLUSIONS
We have developed a method for removal of contaminating DNA in yeast growth media.
Rawsthorne
H
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Department of Bioscience and Biotechnology, Drexel University, Philadelphia, PA, USA.
Phister
T G
TG
D001386:Azides / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D015003:Yeasts / Q000254:growth & development*
D001386:Azides; D003470:Culture Media; D004271:DNA, Fungal; C014535:8-azidoethidium