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Recombinant probiotics
Surá, Tereza ; Rittich, Bohuslav (referee) ; Španová, Alena (advisor)
Theoretical part of this thesis focuses on present state and research of recombinant probiotics and their use in food industry and health care. It also concentrates on their beneficial effects on the health of individuals. The experimental part focuses on the identification of specific bacterial strain in a probiotics food supplements. The DNA was isolated from these products by use of magnetic microparticles and obtained DNA was subsequently analysed through polymerase chain reaction.
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Optimalisation of a new micromethod of DNA isolation from foods
Surá, Tereza ; Rittich, Bohuslav (referee) ; Španová, Alena (advisor)
The thesis were focused on the optimalization of micromethod for isolation of DNA in quality for polymerase chain reaction (PCR) using magnetic microparticles from plant food products. There were chosen a red beetroot (fresh, frozen, dried and sterilized) for the analysis and food products containing red beetroot. Different approaches of processing of homogenates were compared and optimized. The homogenates were prepared in lysis buffer with cetyltrimethylammonium bromide (CTAB) with different amounts of NaCl with or without addition of organic extraction agents chloroform-octanol and isopropanol. Microisolation of DNA was performed using magnetic particles P(GMA). The concentration of NaCl and polyethylene glycol (PEG) 6000 in separation mixtures was tested. The influence on quantity and purity of isolated DNA was compared and the optimum amounts of NaCl in CTAB buffer and optimal concentration of PEG 6000 in separation mixtures were compared. The optimized separation mixture for the DNA isolation from red beetroot was applied to food products containing red beetroot. Amplifiability of DNA was tested in conventional PCR using specific primers for plant DNA. PCR products of length 700 bp were detected by agarose gel electrophoresis.
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Optimalisation of a new micromethod of DNA isolation from foods
Surá, Tereza ; Rittich, Bohuslav (referee) ; Španová, Alena (advisor)
The thesis were focused on the optimalization of micromethod for isolation of DNA in quality for polymerase chain reaction (PCR) using magnetic microparticles from plant food products. There were chosen a red beetroot (fresh, frozen, dried and sterilized) for the analysis and food products containing red beetroot. Different approaches of processing of homogenates were compared and optimized. The homogenates were prepared in lysis buffer with cetyltrimethylammonium bromide (CTAB) with different amounts of NaCl with or without addition of organic extraction agents chloroform-octanol and isopropanol. Microisolation of DNA was performed using magnetic particles P(GMA). The concentration of NaCl and polyethylene glycol (PEG) 6000 in separation mixtures was tested. The influence on quantity and purity of isolated DNA was compared and the optimum amounts of NaCl in CTAB buffer and optimal concentration of PEG 6000 in separation mixtures were compared. The optimized separation mixture for the DNA isolation from red beetroot was applied to food products containing red beetroot. Amplifiability of DNA was tested in conventional PCR using specific primers for plant DNA. PCR products of length 700 bp were detected by agarose gel electrophoresis.
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Recombinant probiotics
Surá, Tereza ; Rittich, Bohuslav (referee) ; Španová, Alena (advisor)
Theoretical part of this thesis focuses on present state and research of recombinant probiotics and their use in food industry and health care. It also concentrates on their beneficial effects on the health of individuals. The experimental part focuses on the identification of specific bacterial strain in a probiotics food supplements. The DNA was isolated from these products by use of magnetic microparticles and obtained DNA was subsequently analysed through polymerase chain reaction.
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