Open AccessRapid identification of pickle yeasts by fluorescent PCR and microtemperature‐gradient gel electrophoresis
Author(s) -
Tominaga Tatsuya
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2004.tb09735.x
Subject(s) - yeast , temperature gradient gel electrophoresis , primer (cosmetics) , biology , 16s ribosomal rna , isolation (microbiology) , fluorescence , chromatography , food science , chemistry , biochemistry , microbiology and biotechnology , bacteria , genetics , physics , quantum mechanics , organic chemistry
Monitoring the yeast populations within pickle soaking fluid is imperative for ensuring optimum taste, but these analyses have proven time‐consuming and expensive, limiting their industrial application. Here, yeasts were identified in the soaking fluid from Japanese radish pickles using fluorescent PCR amplification of the variable D1/D2 region of the 26S rDNA, followed by analysis with microtemperature‐gradient gel electrophoresis (μ‐TGGE). This smaller version of the normal TGGE apparatus is capable of analyzing samples 10‐ to 20‐fold faster without sacrificing data quality. Each primer set was labeled with a different fluorescent dye, allowing easy isolation of the various PCR products and identification of the bands corresponding to the various yeasts. The results indicate that fluorescent PCR and μ‐TGGE may be a useful new method for rapid, easy monitoring of yeast flora in various food industries. This new method can be used on a daily basis to provide overviews of yeast flora during pickle production, allowing producers to quickly grasp pickle readiness at a single glance.