Isolates of yeasts and bacteria were identified by a combination of
phenotypic and molecular methods. Briefly, the Polymerase Chain Reaction
(PCR) was performed to amplify the 5.8S-Internally Transcribed
Spacer rRNA gene region of yeast isolates and the 16S rRNA gene of
bacterial isolates. DNA amplifications were done with the primers ITS1
and ITS4 for yeasts, 27F and 1495R for LAB and 16Sd and 16Sr for acetic
acid bacteria. Following amplifications, PCR products were used for
identification of yeasts and bacteria to species by a combination of restriction
fragment length polymorphism (RFLP) and sequence analyses.
Growth at 37 °C and ascospore production were used as additional tests
to differentiate the yeast Hanseniaspora guilliermondii from other
species (Cadez and Smith, 2011). The procedures and conditions of
DNA extraction, PCR, RFLP and sequence analyses, including the use of
reference cultures, were done as described by Ho et al. (2014).
Isolates of yeasts and bacteria were identified by a combination ofphenotypic and molecular methods. Briefly, the Polymerase Chain Reaction(PCR) was performed to amplify the 5.8S-Internally TranscribedSpacer rRNA gene region of yeast isolates and the 16S rRNA gene ofbacterial isolates. DNA amplifications were done with the primers ITS1and ITS4 for yeasts, 27F and 1495R for LAB and 16Sd and 16Sr for aceticacid bacteria. Following amplifications, PCR products were used foridentification of yeasts and bacteria to species by a combination of restrictionfragment length polymorphism (RFLP) and sequence analyses.Growth at 37 °C and ascospore production were used as additional teststo differentiate the yeast Hanseniaspora guilliermondii from otherspecies (Cadez and Smith, 2011). The procedures and conditions ofDNA extraction, PCR, RFLP and sequence analyses, including the use ofreference cultures, were done as described by Ho et al. (2014).
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