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The influence of storage time on authentication of plant-based foods
Trödlerová, Martina ; Šislerová, Lucie (referee) ; Fialová, Lenka (advisor)
Fruit baby foods and purees belong among adulterated food, due to the prices of materials for their production. More expensive ingredients are replaced by cheaper, lower quality ones. This bachelor thesis deals with influence of storage time on the determination of samples authenticity The theoretical part of the thesis deals with the chemical composition of fruit (especially apples and peaches), production technology and methods of determining authenticity. In the experimental part, model samples were prepared and DNA was isolated from them. Then, using two different primer pairs, this DNA was amplified by real-time PCR and a specific product was detected by analyzing melting curves. On gel electrophoresis, it was possible to confirm the presence of these specific products. The last method was HPLC, where phenolic substances were analyzed.
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Detection of the HLA-B27 allele using a commercial kit
VOLKOVÁ, Simona
The main topic of the presented work is the detection of the HLA-B27 allele using a commercially available kit from the company BAG HEALTH CARE. The HLA-B27 molecule has been linked to some diseases. The best known is the association with ankylosing spondylitis or Bechterew's disease. Ankylosing spondylitis is a lifelong disease, manifested by an inflammatory reaction mainly at the site of the vertebrae of the spine and large joints such as the hips, shoulders and knees. Thanks to the association, the detection of the HLA-B27 allele can be used as a molecular marker for early diagnosis, mitigation and improvement of the course of treatment of ankylosing spondylitis. The theoretical part of the thesis specifies ankylosing spondylitis and the main histocompatibility complex with emphasis on the key molecule HLA-B27. The practical part of the bachelor thesis is focused on sample collection, DNA isolation and DNA amplification. This is followed by the laboratory detection of the HLA-B27 allele itself using a commercial kit using gel electrophoresis to visualize DNA fragments. Thus, 20 anonymized DNA samples from volunteers were examined. Of the 20 samples examined, positivity for the HLA-B27 allele was demonstrated in 6 samples of volunteers who had different clinical manifestations of Bechterew's disease.
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Comparison of gel and capillary electrophoresis of proteins
Krbec, Michal ; Franěk, Tomáš (advisor) ; Kotaška, Karel (referee)
Electrophoresis of serum proteins is one of the methods used in laboratory diagnosis of pathological conditions, but also for research purposes. This is a one-step method, which involves isolation of individual proteins from a mixture (mostly blood proteins) in the electric field in the presence of a suitable buffer and a suitable carrier based on their different mobility, dependent on their surface charge and molecular weight. Originally the carrieer used to be a paper, later cellulose acetate membrane and now it is almost exclusively agarose (purified agar with low endoosmosis) or polyacrylamide (PAA). My task in this work was to analyze a serie of samples by gel electrophoresis and subsequently analyze the same serie by capillary electrophoresis. Results obtained by both methods I worked numerically, compared with each other and statistically evaluated. The output of my work is a comparison of gel and capillary electrophoresis, evaluation of the advantages, disadvantages and especially the differences between these two methods.
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Effect of crowding agents on DNA-surfactant interaction
Sovová, Šárka ; Hurčíková, Andrea (referee) ; Pekař, Miloslav (advisor)
Tato diplomová práce se zabývá vlivem zaplňovacích činidel na interakce v systému DNA-tenzid. DNA o velikosti 4017 párů bází byla připravena polymerázovou řetězovou reakcí, jako templát byl použit plasmid pSB-E1g. Polyetylen glykol (PEG) byl použit jako zaplňovací činidlo a jeho vliv na DNA-tenzid interakce byl zkoumán experimenty založenými na fluorescenci a gelové elektroforéze. Také byl studován vliv iontové síly za použití NaBr na interakce DNA-tenzid za použití zaplňovacího činidla. Data byla vyhodnocena a evaluována v této práci. V úvahu byl brán i možný vliv polyetylen glykolu na kritickou micelarní koncentraci (CMC) tenzidu, bylo provedeno měření CMC pomocí ultrazvuku s vysokým rozlišením, avšak nebyl zjištěn žádný značný vliv zaplňovacího činidla na CMC tenzidu. Část této práce bude zahrnuta v publikaci s anglickým názvem Combined role of macromolecular crowding and cationic surfactant in efficient DNA condensation.
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Comparison of gel and capillary electrophoresis of proteins
Krbec, Michal ; Franěk, Tomáš (advisor) ; Kotaška, Karel (referee)
Electrophoresis of serum proteins is one of the methods used in laboratory diagnosis of pathological conditions, but also for research purposes. This is a one-step method, which involves isolation of individual proteins from a mixture (mostly blood proteins) in the electric field in the presence of a suitable buffer and a suitable carrier based on their different mobility, dependent on their surface charge and molecular weight. Originally the carrieer used to be a paper, later cellulose acetate membrane and now it is almost exclusively agarose (purified agar with low endoosmosis) or polyacrylamide (PAA). My task in this work was to analyze a serie of samples by gel electrophoresis and subsequently analyze the same serie by capillary electrophoresis. Results obtained by both methods I worked numerically, compared with each other and statistically evaluated. The output of my work is a comparison of gel and capillary electrophoresis, evaluation of the advantages, disadvantages and especially the differences between these two methods.
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The interactions of hyaluronan and DNA
Sklenářová, Renáta ; Krouská, Jitka (referee) ; Hurčíková, Andrea (advisor)
This diploma thesis deals with the study of possible interactions between hyaluronan (HA) and plasmid DNA (pDNA). Plasmid DNA was isolated from E. coli JM109 (pUC19) and resuspended in TE buffer as well as high molecular weight hyaluronan. Individual samples of pDNA, HA and pDNA-HA were characterized by gel electrophoresis, CD spectroscopy and high resolution ultrasonic spectroscopy. Agarose gel electrophoresis examined the effect of the addition of hyaluronan to plasmid DNA on the migration of samples to the positive electrode. Structural changes in pDNA-HA samples were examined using CD spectroscopy. Individual CD spectra describes the dependence of the difference in absorption coefficients for left-hand and right-handed elliptic polarized light at wavelength. High resolution ultrasonic spectroscopy has been used to study interactions. It is an analytical method based on ultrasonic velocity and attenuation. We classify this technique as a non-destructive method because the passing waves do not affect the structure of the analyzed sample.
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Preprocessing of 1D gel electrophoresis image
Hlavatý, Matej ; Harabiš, Vratislav (referee) ; Vítek, Martin (advisor)
The aim of this project is an automatic analysis of 1D gel electrophoresis results. Basic principles of gel electrophoresis and types of errors and signal noise influencing the result are studied. The proposed algorithm serves for automatic detection of lanes in the image. Summation projection on the horizontal axis is created from black-and-white digital pictures of electrophoresis. Local minima of the profile are located after smoothing the profile. In the location of these minima borders between lanes are established.
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Automatic improvements of images from 1D gel electrophoresis
Kovář, Martin ; Vítek, Martin (referee) ; Škutková, Helena (advisor)
In this bachelor’s thesis are explained the basic principles of electrophoresis and its modalities with focusing on 1D gel electrophoresis. It describes analysis of an electrophoreogram and causes of its possible distortion, and states specifications of applications of the method in microbiology, genomics and proteomics. The practical part presents development, optimization and outputs of a programme for automatic electrophoreogram analysis, which was created in Matlab environment. The analysis contains lane and band detection and computation of samples’ molecular weight. The ending of the thesis is constituted by evaluating efficiency of detection and accuracy of weight computation.
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Plasmid DNA interactions with lanthanoide compounds
Gőghová, Sabína ; Španová, Alena (referee) ; Rittich, Bohuslav (advisor)
In the theoretical part of bachelor thesis, literature was summarised on the isolation and purification of plasmid DNA, the application of lanthanides as non enzymatic nucleases and practical importance of lanthanide compounds in medicine. In the experimental part of the thesis, plasmid DNA pUC19 was isolated by the method of alkaline lysis. Isolated plasmid DNA was cleaved by neodymium trichloride at 70 °C in different intervals. Gel electrophoresis was used for evaluation of experiments.
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Plasmid DNAs interactions with lanthanoide compounds
Budko, Kateryna ; Horák, Daniel (referee) ; Rittich, Bohuslav (advisor)
Recently much attention is given to lanthanides and their complexes as excellent catalysts for cleavage of nucleic acids. The thesis has been focused on the cleavage of plasmid and bacterial DNA by ions Nd3+ and Y3+ and by different carriers containing the lanthanide compounds. The creation of single-stranded nicks and double-stranded ones in the plasmid DNA molecules was studied by agarose gel electrophoresis. Verification of the cleavage of bacterial DNA was made by polymerase chain reaction using primers specific for the domain Bacteria and genus and species-specific primers. The results will be used in the development of the method that will allow perfect carriers`s coverage verification with the magnetic perovskit nucleus and other carriers with the lanthanide compounds.
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