Future Methods

The use of PCR in molecular microbiology has increased to the point where it is now accepted as the golden standard for detecting nucleic acids from a number of origins and has become an essential tool in the research laboratory. Real-time PCR has engendered wider acceptance of PCR, due to its improved rapidity, sensitivity, reproducibility, and the reduced risk of carry-over contamination (Mackay et al. 2002). Traditional detection of amplified DNA relies upon electrophoresis of the nucleic acids in the presence of ethidium bromide and visual observation after irradiation by ultraviolet light. Southern blot detection of the amplicon using hybridization with a labeled oligonucleotide probe is also time consuming. The detection of the amplicon and the possibility of visualizing it as the amplification progresses is the foundation of real-time PCR. The monitoring of the accumulation of the amplicon in real time has been made possible by the labeling of primers with fluorogenic molecules. Due to the numerous advantages, these methods probably are an alternative to the techniques described previously, not only for the detection and identification of spoilage yeast or for wine yeast monitoring during the alcoholic fermentation. Thus, this technology has also been applied in studies of yeast gene expression during the alcoholic fermentation.

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