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Yeasts and wine
Palíková, Petra ; Vadkertiová, Renata (referee) ; Vránová, Dana (advisor)
This thesis deals with isolation and identification wine yeasts from grapes and must. For analysis was used white wine Sauvignon that was grown and producing after needs ecological agriculture. Remove samples were processed in laboratory and by the help of dilution method were obtained pure culture isolated yeasts. In the following step, by the application of commercial kit UltraCleanTM Microbial DNA Isolation Kit we were able to isolated individual DNA that it was used to the next analysis. Isolated DNA was amplification by PCR method with ITS1 and ITS4 primers. PCR products were detected on agarose gel. Amplification samples were chopped five restriction endonucleases: HaeIII, HinfI, TaqaI, AluI and MseI. Chopped DNA was detected by the same way as PCR products and it was compared with restriction patterns of collection yeasts. In the next step it was compared genetic similarity of isolated yeasts by using BioNumerics software. As a criterion it was used Pearson coefficients and UPGMA clastering analysis. The result is dedrogram of genetics similarity isolated yeasts.
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Preprocessing of 1D gel electrophoresis image
Svozilová, Veronika ; Škutková, Helena (referee) ; Vítek, Martin (advisor)
The aim of this work is to propose an algorithm in MATLAB, allowing image processing and evaluation of one-dimensional gel electrophoresis. At first describes the detection of samples based on the horizontal segmentation of electrophoresis gel image, followed by detection of the minimum intensity values of the image, a vertical segmentation and determination of the sample boundaries in the image. The final phase involves testing of the method on publicly available images and evaluation of the results.
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Identification of PHA producing bacteria employing molecular techniques
Gajdová, Barbora ; Obručová,, Hana (referee) ; Obruča, Stanislav (advisor)
This diploma thesis deals with identification of bacteria which are capable of producing polyhydroxyalkanoates (PHAs). Work included testing variety of genera including Pseudomonas, Lactobacillus, Bifidobacterium, thermophilic cultures and samples gathered from natural sources. Bacteria were investigated by molecular technique polymerase chain reaction – PCR. An amplification of the PHA synthase gene (phaC) was analyzed. In the first reaction phaC and 16S rRNA genes were tested at the same time. 16S rRNA gene is used as control for bacterial DNA and as an identification tool for natural source samples. This multiplex PCR used multiple primers in PCR mix. Second reaction search for amplicon specific for catalysing biosynthesis mcl-PHA (phaC1). The presence of the PHA synthase gene was verified in 11 samples which were Bifidobacterium breve CCM 7825T, Lactobacillus rhamnosus CCM 1825T, Lactobacillus zeae CCM 7069T, Lactobacillus delbrueckii subsp. bulgaricus CCM 7190T, Lactobacillus plantarum CCM 7039T, Pseudomonas gessardii, Pseudomonas fulva, Arthrobacter protophormiae, Curtobacterium flaccumfaciens, Mycobacterium neoaurum and Staphylococcus lentus.
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Compensation of geometric distortion of electrophoretic gel image
Dvořáček, Tomáš ; Vítek, Martin (referee) ; Škutková, Helena (advisor)
This master thesis is engaged in problematics of creation and compensation of geometric distortions in 1D agarose electrophoresis. This master thesis analyze the problematics of cause of these distortions and summarize the theory needed for compensation of these distortions. Based on acquired theory and created electrophoretic phantoms, the master thesis contains several suggestions for compensation of incurred distortions. These suggestions are recreated into functions, which are connected into a functional user interface for gel image analysis and geometric distortions compensation.
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Evaluation of basic characteristics of thermal water with separation methods
Skálová, Lucie ; Mravcová, Ludmila (referee) ; Vávrová, Milada (advisor)
The presence of inorganic ions is one of the criteria for thermal water assessing. The most important anions that affect the water quality and allow their use for therapeutic purposes, include sulfates, chlorides, nitrates and bicarbonates. Ions (as sodium, potassium, calcium and magnesium) are the significant cations. The ion chromatography is used for separation of these substances and also some methods based on electrophoretic migration of ions in an electric field. The electromigration method of isotachophoresis was chosen for determination of selected ions in water samples collected from thermal boreholes in Pasohlávky.
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Analysis of low-molecular proteins in barley by the SDS-PAGE method during malting
Myslivcová, Pavla ; Svoboda, Zdeněk (referee) ; Mikulíková, Renata (advisor)
This diploma thesis deals with the analysis of low molecular weight proteins in barley during malting by SDS-PAGE method. Attention was paid to PR proteins, specifically LTP proteins and thionins, considered to be connected with gushing effect. Barley samples, malt intermediates and malt samples taken in 10 consecutive days to cover the whole malting process were used for the experiment. In total, 5 samplings were used for the analysis. Proteins extracted from the samples were separated by SDS-PAGE using a Tris-tricine buffer system. The protein lines of LTP proteins and thionins were identified in the resulting gels. The relative optical density values of the selected protein bands were obtained to assess the effect of malting on these proteins. A similar pattern of change in the content of mentioned low molecular weight proteins during the malting process was observed. This was confirmed by finding a statistically significant positive correlation between the relative optical density values of LTP proteins and thionins. Furthermore, the relationship between the low molecular weight protein content and the gushing potential and the microbiological contamination of the samples was investigated, but was not confirmed.
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Application for the Data Processing in the Area of Genome Engineering
Brychta, Jan ; Burgetová, Ivana (referee) ; Očenášek, Pavel (advisor)
This masters thesis has a few objectives. One of them is to acquaint with the problems of genome engineering, especially with fragmentation of DNA, the macromolecule DNA, the methods for purification and separation of the nucleic acids, the enzymes used for modification of these acids, amplification and get to know with cluster and gradient analysis as well. The next aim is to peruse the existed application and compare it to the layout of the proposed application, that is the third aim. The last one from the objectives is the implementation and the report how was the application tested by the real data. The results will be discussed as well as the possibilities of the further extension.
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Changes of gluten proteins during beer processing
Porubiaková, Otília ; Mikulíková, Renata (referee) ; Márová, Ivana (advisor)
The aim of thesis was monitoring of changes in the content of gluten proteins in the biotechnological process of beer production. During the production process of wheat and barley beer, the samples were collected and analysed using the electrophoresis and immunoassay method. The results of the analyses were compared with commercial Czech beers. The theoretical part contains description and composition of gluten proteins, malt and beer technology, the changes that occur in this process, and methods of gluten proteins analysis. The experimental part contains procedures for laboratory production of barley and wheat beer and analyses of gluten proteins. To identify the individual gluteal protein fraction acid and SDS electrophoresis methods were used. For quantification, enzyme immunoassay was used and evaluated spectrophotometrically. The identification of the gluten‘s fractions by electrophoretic methods has been shown to be less specific for samples with lower content of gluten proteins and for barley specimens. A decrease in the concentration of gliadins and glutenins in the beer production process was demonstrated. A significant change was found during wort production with 98% decrease of gluten content compared to the feedstock and during the fermentation, when the gluten concentration dropped below 10 mg/kg. This value is acceptable from the legislation for products labelled „gluten-free“.
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Dynamic spot analysis in the 2D electrophoresis gels images
Polášková, Lenka ; Provazník, Ivo (referee) ; Nedvěd, Jiří (advisor)
Práce shrnuje faktory a parametry, které ovlivňují výsledky 2D elektroforézy, se zaměřením na zpracování obrazu jako jeden ze způsobů snížení nesprávné interpretace jejích výstupů. Proces zpracování obrazu využívá jako zdroj dat především obrazů z opakovaných provedení téhož pokusu, neboli víceplik. Pomocí analýzy obrazů víceplik je možno pozorovat nebo korigovat změny jednoho pokusu a také porovnávat je s výstupy jiných pokusů. Cílem práce je poskytnout podporu specialistovi, který má na starosti popsat vlastnosti struktur nacházejících se v elektroforetických obrazech.
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