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Comparison of different types of magnetic carriers for DNA microisolation from foods
Koplík, Jerguš ; Lunerová,, Jana (referee) ; Kovařík, Aleš (advisor)
Micro-isolation of PCR-ready from fresh and dried legumes seeds and food products containing legumes (hummus) was tested. For optimization process magnetic microparticles PGMAox was used. Optimum weight plants and food material was 200 mg. For isolation of DNA from fresh legumes, mixture containing 500 L of CTAB extraction buffer, 1 L -mercaptoethanol and 500 L chloroform-octanol was used. For isolation of DNA from dried legumes seeds and food products volume of CTAB extraction buffer was increased to 1 000 L. To achieve higher purity of DNA some prepared homogenates was precipitate by isopropylalcohol. The optimized process of DNA isolation was used to prepare a homogenate from food products which DNA was isolated by different types of magnetic carriers. For comparison, non-porous magnetic carriers poly(glycidyl methacrylate) PGMAox, poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) P(HEMA-co-GMA)ox covered by carboxyl groups and porous fully crosslinked microparticles poly(styrene-co-divinylbenzene) HPS-B-M-22-NH2, magnetic porous glass MPG and nanoparticles of iron oxides covered by poly(L-lysine) PLL were used. In average the highest concentration and the best purity of DNA was isolated by magnetic carriers P(HEMA-co-GMA)ox a PGMAox.
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Comparison of different types of magnetic carriers for DNA microisolation from foods
Koplík, Jerguš ; Lunerová,, Jana (referee) ; Kovařík, Aleš (advisor)
Micro-isolation of PCR-ready from fresh and dried legumes seeds and food products containing legumes (hummus) was tested. For optimization process magnetic microparticles PGMAox was used. Optimum weight plants and food material was 200 mg. For isolation of DNA from fresh legumes, mixture containing 500 L of CTAB extraction buffer, 1 L -mercaptoethanol and 500 L chloroform-octanol was used. For isolation of DNA from dried legumes seeds and food products volume of CTAB extraction buffer was increased to 1 000 L. To achieve higher purity of DNA some prepared homogenates was precipitate by isopropylalcohol. The optimized process of DNA isolation was used to prepare a homogenate from food products which DNA was isolated by different types of magnetic carriers. For comparison, non-porous magnetic carriers poly(glycidyl methacrylate) PGMAox, poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) P(HEMA-co-GMA)ox covered by carboxyl groups and porous fully crosslinked microparticles poly(styrene-co-divinylbenzene) HPS-B-M-22-NH2, magnetic porous glass MPG and nanoparticles of iron oxides covered by poly(L-lysine) PLL were used. In average the highest concentration and the best purity of DNA was isolated by magnetic carriers P(HEMA-co-GMA)ox a PGMAox.
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Properties of probiotics used in food and their safety
Koplík, Jerguš ; Němcová, Andrea (referee) ; Trachtová, Štěpánka (advisor)
The thesis deals with the criteria and the properties which the microorganisms must fulfill to be included among the probiotics. After it deals with the assessing of the safety of probiotic strains used in the food industry nowadays and their potential negative influence on the food quality and the human health. Then the thesis defines the kind of cheese Bryndza like the potential probiotic product. It focuses on the microbial composition, the process passing during the production and the general qualities of this kind of regional food. The aim was to analyse microflora of cheese Bryndza and that´s why PCR was carried out in the real time by using primers, which are typical for domain Bacteria, total yeasts and genus Lactobacilli. Three different samples of cheese Bryndza were analysed. DNA was isolated by two types of magnetic carriers. To find out the particularities of these products the analysis of the melting curve was carried out and its results were verified by the gel electrophoresis. The existence of bacterial DNA and lactobacilli was proved in two samples. In the third one there was lower, even no concentration. The existence of yeasts was not proved only in one of the samples.
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