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Evaluation of a micromethod for isolation of DNA from plant leaf, fruit and fruit products
Balažovičová, Nikola ; Rittich, Bohuslav (oponent) ; Kovařík, Aleš (vedoucí práce)
The thesis has been focused on testing of micromethod of DNA isolation from leaves, fruits and fruit products. Jams were selected for the analysis of plant DNA in technologically processed foods. Plant leaves, fruits, and jams were homogenized using plastic copist in a lysis buffer containing 2% cetyltrimethylammonium bromide (CTAB) with 2.5M sodium chloride (NaCl). Microisolation of plant DNA was performed using poly(hydroxyethylmethacrylate-co-glycidylmethacrylate) – P(HEMA-co-GMA)microparticles. Isolated the DNA concentration and purity were assessed by UV light aborbance using a spectrophotometer. After that, amplification of the DNA was tested in PCR. Primers specific for plant ribosomal DNA: 18S_for a 5,8S_rev (PCR product - 700bp), 26S_for a 26S_rev (PCR product - 220 bp), 18S_for a 18¬S_rev (PCR product - 263 bp) were used. The PCR conditions were optimized and the effect of the amplicon length on its detection was followed. PCR products were detected by agarose gel electrophoresis. It was shown that DNA isolated from almost all of leaves using magnetic particles was in PCR-ready quality in contrary to the fruits. DNA amplified in PCR with primers giving short PCR products was isolated from almost all tested jams. The method must be optimalised, yet.
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Bezpečnost probiotik používaných v potravinách
Balažovičová, Nikola ; Rittich, Bohuslav (oponent) ; Španová, Alena (vedoucí práce)
V teoretickej časti práce je pozornosť zameraná na definíciu probiotík, na ich zdravotné výhody, na probiotické potraviny, na kritéria bezpečnosti probiotík, vrátane metód identifikácie probiotických baktérií. V praktickej časti práce bola pozornosť zameraná na identifikáciu probiotických baktérií obsiahnutých v mliečnom výrobku Actimel. DNA v kvalite vhodnej pre PCR bola izolovaná pomocou magnetických mikročastíc P(HEMA-co-GMA). Identifikácia bola prevádzaná metódou polymerázovej reťazovej reakcie. Gélovou elektroforézou produktu sa potvrdila prítomnosť bakteriálnej DNA pre doménu Bacteria, rod Lactobacillus a druh Lactobacillus casei. Špecifické produkty PCR o veľkostiach 466 bp (doména Bacteria), 250 bp (rod Lactobacillus) a 136 bp (druh Lactobacillus casei) boli detekované po amplifikácii DNA izolovanej z probiotického mliečneho výrobku.
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Evaluation of a micromethod for isolation of DNA from plant leaf, fruit and fruit products
Balažovičová, Nikola ; Rittich, Bohuslav (oponent) ; Kovařík, Aleš (vedoucí práce)
The thesis has been focused on testing of micromethod of DNA isolation from leaves, fruits and fruit products. Jams were selected for the analysis of plant DNA in technologically processed foods. Plant leaves, fruits, and jams were homogenized using plastic copist in a lysis buffer containing 2% cetyltrimethylammonium bromide (CTAB) with 2.5M sodium chloride (NaCl). Microisolation of plant DNA was performed using poly(hydroxyethylmethacrylate-co-glycidylmethacrylate) – P(HEMA-co-GMA)microparticles. Isolated the DNA concentration and purity were assessed by UV light aborbance using a spectrophotometer. After that, amplification of the DNA was tested in PCR. Primers specific for plant ribosomal DNA: 18S_for a 5,8S_rev (PCR product - 700bp), 26S_for a 26S_rev (PCR product - 220 bp), 18S_for a 18¬S_rev (PCR product - 263 bp) were used. The PCR conditions were optimized and the effect of the amplicon length on its detection was followed. PCR products were detected by agarose gel electrophoresis. It was shown that DNA isolated from almost all of leaves using magnetic particles was in PCR-ready quality in contrary to the fruits. DNA amplified in PCR with primers giving short PCR products was isolated from almost all tested jams. The method must be optimalised, yet.
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Bezpečnost probiotik používaných v potravinách
Balažovičová, Nikola ; Rittich, Bohuslav (oponent) ; Španová, Alena (vedoucí práce)
V teoretickej časti práce je pozornosť zameraná na definíciu probiotík, na ich zdravotné výhody, na probiotické potraviny, na kritéria bezpečnosti probiotík, vrátane metód identifikácie probiotických baktérií. V praktickej časti práce bola pozornosť zameraná na identifikáciu probiotických baktérií obsiahnutých v mliečnom výrobku Actimel. DNA v kvalite vhodnej pre PCR bola izolovaná pomocou magnetických mikročastíc P(HEMA-co-GMA). Identifikácia bola prevádzaná metódou polymerázovej reťazovej reakcie. Gélovou elektroforézou produktu sa potvrdila prítomnosť bakteriálnej DNA pre doménu Bacteria, rod Lactobacillus a druh Lactobacillus casei. Špecifické produkty PCR o veľkostiach 466 bp (doména Bacteria), 250 bp (rod Lactobacillus) a 136 bp (druh Lactobacillus casei) boli detekované po amplifikácii DNA izolovanej z probiotického mliečneho výrobku.
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