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Plasmide DNA isolation from bacteria and transfection to HEK293 cell line
Bačovská, Kristýna ; Chmelíková, Larisa (referee) ; Svoboda, Ondřej (advisor)
Plasmid DNA is commonly used in fields of molecular biology and genetic engineering. This work deals with methods of DNA isolation and topics related to this procedure. In the experimental part of the work, phenol-chloroform extraction is used. First of all, plasmids Channelrhodopsin-2, ASAP1 and Kir 2.1 were amplified in bacterial strain DH5. 22 isolations were accomplished and the yield was validated using gel electrophoresis and transfection to HEK293 cell line. The most successful isolation was the isolation of plasmid ASAP1, the overall percentage success rate was 30 %.
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Web Information System Design Proposal
Svoboda, Ondřej ; Koutný, Miroslav (referee) ; Dydowicz, Petr (advisor)
This bachelor’s thesis deals with evaluation and design of a website, where will also be designed information website system (further IS). It will have a form of a calendar and it should help the firm to better time planning, organization of work, guidance of employees and echo checking of information. In IS there will be modules for evidence of workers, customers, orders, services, costs and particular jobs. IS design will be like that so there is an opportunity to add extending modules.
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Friction and air feeders for paper sheets
Svoboda, Ondřej ; Košťál, David (referee) ; Brandejs, Jan (advisor)
The thesis is focused on summarizing the evidence on the issue of paper sheets feeding at the final processing of documents. The thesis includes dividing of paper feeders, their characterization, and description of friction and air feeders, which are most widespread. As a model for the description methods of sheets feeding were used machinery CFM 700 and GPM 540A of Cyklos company. The thesis also compares friction and air feeding from the constructional point of view. The conclusion focuses on the new methods in the paper feeding in future.
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Electrophysiological characterization of Kir2.1 membrane channel
Měsíčková, Klára ; Chmelíková, Larisa (referee) ; Svoboda, Ondřej (advisor)
The topic of this thesis is electrophysiological characterization of Kir2.1 membrane channel. Inward rectifier potassium channel Kir2.1 is located in muscular, heart and nerve cells and its dysfunction causes various diseases. Practical part of this stage is focused on cultivation of the HEK293T cell line that is used to transfection of the plasmid Kir2.1 and subsequent measurement of the ionic current through the electrophysiological method patch-clamp in whole-cell mode.
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Analysis of ZED stereocamera in outdoor environment
Svoboda, Ondřej ; Věchet, Stanislav (referee) ; Krejsa, Jiří (advisor)
The Master thesis is focused on analyzing stereo camera ZED in the outdoor environment. There is compared ZEDfu visual odometry with commonly used methods like GPS or wheel odometry. Moreover, the thesis includes analyses of SLAM in the changeable outdoor environment, too. The simultaneous mapping and localization in RTAB-Map were processed separately with SIFT and BRISK descriptors. The aim of this master thesis is to analyze the behaviour ZED camera in the outdoor environment for future implementation in mobile robotics.
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Information System for a School
Svoboda, Ondřej ; Fiala, Jiří (referee) ; Částek, Petr (advisor)
This bachelor's thesis describes designing and creation of information system for Higher school of education, which will improve communication between students and lectors. Final application is written in PHP scripting language and uses MySQL database and AJAX technology. During implementation object-oriented programming paradigm was used.
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Plasmide DNA isolation from bacteria and transfection to HEK293 cell line
Skala, Kateřina ; Fohlerová, Zdenka (referee) ; Svoboda, Ondřej (advisor)
DNA isolation is one of the basic methods in molecular biology. There are several methods of DNA amplification and isolation. In this paper phenol-chloroform extraction of three plasmid types - Channelrhodopsin-2, ASAP1 and Kir 2.1 is used. Six plasmids were isolated in total. These plasmids are then validated using gel electrophoresis. Successfully isolated plasmids are then transfected to HEK293 cells and images taken on confocal microscope 24, 48 and 72 hours after transfection.
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Project of revitalization part of Kninicky brook
Vyplel, Adam ; Svoboda, Ondřej (referee) ; Šlezingr, Miloslav (advisor)
The diploma thesis is focused on the proposal of systematic revitalization of the selected section of the Knínický brook in the cadastral area of Veverské Knínice. It is a straight, very deep channel due to technical adjustments in the past. The HEC-RAS program verifies the capacity of the channel for the Q100 flow. In this way, it was found that the channel has very high capacity so a new route with Q1 flow capacity was designed. Due to the height conditions, the design also included three boulder chutes. Subsequently, the vegetation accompaniment and bank stands were suitably supplemented.
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3D bioprinting of stem cells and analysis of microscopic images
Kandra, Mário ; Svoboda, Ondřej (referee) ; Kolářová, Jana (advisor)
In this diploma thesis we are discussing about using 3D bioprinting in tissue engineering. We are discribing using biomaterials for construction scaffolder and aplication stem cells in 3D bioprinting. Last section of theoretical part deals with very often used techniques of 3D bioprinting and we are focused on extrusion technique. In the practical part we propose a method for print vasculars structures. We realized prototype of print head, her design and 3D printing of individual parts. To mechanical part we create a control system for printing control. At the end we visualize the organization of the cells using program modules.
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Study of properties of voltage membrane sensor ASAP1 expressed in HEK293 cell line
Sanetrníková, Dominika ; Chmelíková, Larisa (referee) ; Svoboda, Ondřej (advisor)
In the beginning of this thesis is a short introduction into plasmid DNA which is in the form of a vector used in molecular biology. Plasmids can be used in the form of fluorescent probes to measure changes in membrane potential. Into their structure is added a dye called fluorophore. As an important representative of this thesis is a fluorescent probe ASAP1 which contains green fluorescent protein whose response to the membrane potential change is the decrease in the intensity of emitted light. The aim of this thesis was to make chemical transfection of this plasmid into the HEK293 cell line and carry out its characterization. In the work is also described the design of a method for the analysis of the time course of changes in fluorescence depending on the cell membrane depolarisation. In the end of this thesis is also desribed realized experiment including the discussion of aquired results.
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