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Preparation and purification of plasmid DNAs
Němeček, Milan ; Španová, Alena (referee) ; Rittich, Bohuslav (advisor)
With the development of therapeutic methods in medicine, like are DNA vaccines and gene therapy increases demand for new processes for the isolation and purification highly pure plasmid DNA. Most often used methods of purification plasmid DNA are chromatographic methods. In experimental part of this thesis was performed isolation of plasmid pUC-19 DNA plasmid via alkalyne lysis. And purification of plasmid was performed by liquid chromatography and agarose gel electrophoresis.
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Interaction of Lanthanide trichlorides with the construction materials of a purification apparatus
Novotný, Radoslav ; Zmrzlý, Martin (referee) ; Richtera, Lukáš (advisor)
Lanthanide halides LnX3 are the base material for preparation a different compounds, but also the pure metals. Our faculty is used for the syntesis organometallic catalyst. This compounds are very sensitive to the presence of moisture and oxygen. Lanthanide halides are commercially available in anhydrous or hydrated form. This thesis is devided eight chapters and deals with matherial of the apparatus, which is designed to sublimation of anhydrous lanthanide halides. Initial chapters described the properties and preparation this substances. Following are on the sublimation and corrosion materials. In the last chapter is the skeleton sketch, layout and a availability parts of construction. The aim of this work was to create theoretical basis of construction aparatus and commissioning.
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Focused eletron beam induced deposition
Juřík, Karel ; Zmeškal, Oldřich (referee) ; Pospíšil, Jan (advisor)
Within this work, a set of depositions induced by focused electron beam was prepared. The depositions were prepared in the presence of water vapours from trimethyl(methylcyclopentadienyl)platinum(IV) precursor. The dependence of prepared materials purity on beam accelerating voltage and water vapour pressure was measured. The best platinum content was achieved at (27,2 ± 0,4) at. %, with beam accelerating voltage 5 kV, beam current 1600 pA and water vapour pressure 100 Pa. Due to subsequent long-term exposure to light, air humidity and air oxygen, the platinum content was increased to (39,2 ± 2,1) at. %.
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Proposal for a new method of purification of polyether polyols by zeolits.
Hedvigy, Marek ; Čamaj,, Vladimír (referee) ; Krištín, Michal (advisor)
This diploma thesis covers the tracking of final quality parameters of polyether polyol – Slovaprop G-160, which are: amount of sodium and potassium, color, acidity and amount of water. The most important parameter was the amount of sodium and potassium, because the aim this thesis was to achieve satisfactory amount of sodium and potassium after the purification process with use of zeolite (ZeoCem Micro 200) and to improve the effectivity of the purification and filtration process in the production of polyether polyol – Slovaprop G-160. In theoretical part, Diploma thesis deals with the method of production and characteristics of polyether polyols for the production of polyurethane foams. Practical part consists of nine series of experiments in order to get addictions of amount of sodium and potassium to temperature, weight of Pyrofos and amount of zeolite. Based on laboratory test a new way of purification process of polyether polyols was developed. With this new purification process the production time of Slovaprop G-160 will be shorten thanks to removal of hydrolysis from the reaction process and in the end of the process the time needed for drainage is reduced in half.
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Production of polyhydroxyalkanoates from waste substrates and their isolation
Grossová, Marie ; Babák, Libor (referee) ; Márová, Ivana (advisor)
The aim of this work is to study the possibility of microbial production of polyhydroxyalkanoates (PHA). PHA can be used as biodegradable materials. Bacterial strain Cupriavidus necator was used for laboratory production of PHA. This bacterium was cultivated in medium with various precursors to produce copolymers of 3HB with 3HV or 4HB. Another part of the work was aimed at cultivation of C. necator on different waste substrates, especially oils, with the aim to achieve the highest production of polymer. Another large part of the thesis is dedicated to isolation strategies of PHA using enzymes. Commercially used proteases – alcalase and pancreatin – can be used with advantages for digestion of bacterial cells. A number of optimization experiments showed that application of proteases leads to enhancement of PHA purity to about 13%. Purity increase up to 90 % was achieved by adding a surfactant, which promotes the solubility of non-PHA forming polymer. This surfactant increases the purity of 20 % when compared to control. The last part of presented work deals with the use of enzyme solution isolated from Bacillus subtilis medium. Its application to C. necator culture led to the yield of polymer at a purity exceeding 95 %. These results could represent the basis for new isolation strategies, which can lead to more efficient yield of PHA.
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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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Nucleic acids as terapeutic agent
Ráčková, Lucie ; Španová, Alena (referee) ; Rittich, Bohuslav (advisor)
With the development of molecular biology development of oncotherapy proceeds. The major progress of modern medicine is gene therapy. In the gene therapy are two categeories, namely, viral vectors and nonviral vectors which are used mainly. Nonviral vectors include plasmids. Plasmid DNA used in medicine must be perfectly purified. Chromatographic methods are mainly used at present. Research and development deals with other methods for example two-phase aqueous systems and magnetic carriers. In experimental part of this thesis, isolation of pUC 19 plasmid DNA from Escherichia coli JM 109 (pUC 19) cell culture was performed via method of alkaline lysis. Quality of isolated plasmid DNA was verified spectrophotometrically and by agarose gel electrophoresis. Isolated plasmid DNA was purified using three methods: RNA in plasmid DNA was precipitated by lithium chloride, RNA was degraded by immobilized RNase A and plasmid DNA was purified using two-phase aqueous system.
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Analysis of trypsin inhibitor-like cysteine-rich domain-containing peptides (TIL-domain inhibitors) from the tick Ixodes ricinus
PECHOVÁ, Hana
The master thesis deals with the analysis of trypsin inhibitor-like cysteine-rich domain-containing peptides (TIL-domain inhibitors) extracted from salivary glands of a tick Ixodes ricinus. It comprises the initial bioinformatical analysis of TIL-domain containing peptide family from I. ricinus, molecular cloning of the representative protein into expression vector, followed by production of recombinant TIL-domain inhibitor in bacteria. A representative TIL-domain gene is proved for their structure, biochemical properties as well as immunomodulatory and anticoagulation features.
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Guilt, Purification, Forgiveness
Slezáková, Veronika ; Keřkovský, Pavel (advisor) ; Halama, Jindřich (referee)
This bachelor thesis deals with the conceptions of Guilt, Purification and Forgiveness. Based on the study of the writings of Karl Jaspers, Božena Komárková, Martin Buber and Helmut Weber, the thesis comes to an understanding of the issues of guilt, forgiveness and purification in both philosophical and theological terms. Furthermore, this work introduces other issues that relate to the chosen topic, for example, the concepts of Repentance, Penance and Conversion. Thanks to the acquired findings, I want to come to an understanding of the concept of guilt in Bernhard Schlink's novel The Reader. This thesis is concerned with Schlink's comprehension of guilt as well as with the foundation for reconciliation with guilt that Schlink outlines. Powered by TCPDF (www.tcpdf.org)
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