Determination of dynamic characteristics of microbiota in a fermented beverage produced by Brazilian Amerindians using culture-dependent and culture-independent methods

The impact of yeast starter cultures on the microbial communities and volatile compounds in cocoa fermentation and the resulting sensory attributes of chocolate

“…All reactions were performed in a 25 μL solution containing 0.125 U Taq DNA polymerase (Promega, Madison,USA), 5 μL 5 × go taq buffer, 0.2 mM dNTP, 1 μM each primer, 3.0 mM MgCl 2 , and 0.5 μL of extracted DNA. The amplification was performed as previously described (Ramos et al 2010). The amplified products (2 μL) were analyzed by electrophoresis on 1 % agarose gels before DGGE analysis.…”

“…The PCR products were separated on polyacrylamide gels [8 % (w/v) acrylamide:bisacrylamide (37.5:1)] in 1 × TAE buffer with a DCode System apparatus (BioRad Universal DCode Mutation Detection System, Richmond, CA, USA), according to the procedures previously described (Ramos et al 2010). Solutions containing 30-65 % denaturant [100 % denaturant contains 7 M urea and 40 % (v/v) formamide] were used for bacteria, and solutions containing 35-60 % denaturant were used for the yeasts.…”

Characterization of Microbial Community during the Fermentation of Chinese Homemade <i>paocai</i>, a Traditional Fermented Vegetable Food

“…Due to these limitations, the powerful culture-independent 16S ribosomal RNA (rRNA) gene-based PCR technique, such as PCR-DGGE (Nie et al, 2015), real-time PCR (Paul et al, 2015), Amplified Ribosomal DNA Restriction Analysis (ARDRA) (Park et al, 2014), fluorescent in situ hybridization (FISH) (Ding et al, 2014), etc., have been widely applied to investigate the diversity and dynamics of the microbial community in the field of food in recent years. PCR-DGGE, a classical molecular ecological technic, has been shown to be a suitable tool and widely applied to directly reveal and rapidly monitor the microbial community in the process of fermentation (Nie et al, 2015;Ramos et al, 2010). PCR-DGGE has been used to study the microbial community structure of fermented vegetable food.…”

Identification of the potential inhibitors of malolactic fermentation in wines

“…Reaction products were resolved by electrophoresis in 1% agarose gels, and they were visualized using ethidium-bromide staining. The purified PCR fragments were used for PCR-DGGE sequencing with the DCode Universal Mutation-Detection System (BioRad, Richmond, CA, EUA) according to Ramos et al (2010). The denaturation gradient ranged from 30% to 60% (where 100% corresponds to urea 7 M and formamide 40% v/v).…”