|
|
Isolation of DNA from touch traces and its forensic analysis
Hnyková, Tereza ; Korabečná, Marie (advisor) ; Stenzl, Vlastimil (referee)
Touch DNA is still a relatively new discovery in forensic genetics, but it is used very often as evidence in court proceedings. Nevertheless, its definition is not uniform and the source of DNA in touch deposits is not completely explored. This work summarizes all sources of DNA in touch traces found so far. At the same time, it will provide a brief look on the research carried out on these experimentally deposited traces in laboratories and how the information obtained can be applied in real cases in forensic practice. Finally, DNA extraction methods and the best methods discovered so far to isolate DNA from touch traces will be summarized. Key words: Touch DNA, Touch traces, Trace DNA, Forensic genetics, cell-free DNA, Corneocytes, DNA extraction, Casework, Criminology
|
|
|
Development of methodology for detection of circulating nucleic acids in cancer patients
Kobrle, Lukáš ; Souček, Pavel (advisor) ; Valihrach, Lukáš (referee)
Cancer diagnosis represents one of the most common causes of death in the world, also linked to an ever-increasing incidence of even some more difficult-to-treat variants. One of the most common types of cancer in the world is colorectal cancer, which ranks second in terms of mortality. For these reasons, various methods have been developed for an early diagnosis of the disease. One of the modern and non-invasive methods is the so-called liquid biopsy, based on the detection of circulating DNA and RNA from the blood of patients. This method is also suitable for choosing a therapy, monitoring its effectiveness, and observing the relapse of the disease. In this work, a methodical workflow was created for the monitoring of circulating nucleic acids for its further use in a cohort of patients with colorectal cancer. Various commercial kits were used for the isolation of circulating nucleic acids from plasma of healthy controls, among which the best ones, in terms of quality and quantity, was chosen. The threshold for hemolysis detection by qPCR was also determined using samples from healthy donors. Everything was then applied to RNA samples extracted from plasma of cancer patients, from whichna library of short RNA molecules was prepared for subsequent sequencing. After processing the results, the...
|
|
|
The function of telomeres and cell-free DNA in the healthy volunteers and patients with chosen pathological condition
Zinková, Alžběta ; Korabečná, Marie (advisor) ; Vodička, Radek (referee) ; Drábek, Jiří (referee)
More than 70 years have passed since the discovery of cell-free DNA (cfDNA), but the greatest interest in this topic and knowledge has undoubtedly occurred in the last thirty years. It is used mainly in oncology and prenatal diagnostics. While it is routinely used diagnostically in these fields, little is known about its physiological functions in the organism. Our research therefore focuses on understanding this role and in experiments works mainly with samples obtained from healthy individuals. The first study focused on the differences between plasma and serum in healthy individuals. We asked the question whether they differ in cfDNA concentration and telomeric sequences abundance. We found that the serum contains significantly more cfDNA than plasma, on the other hand, plasma is relatively richer in telomeric sequences. In stimulation experiments with THP1 cells, samples cultured with DNase-treated serum (without cfDNA) showed a higher expression of mRNA TNF-α (Tumor necrosis factor α) than samples untreated. The same trend was observed when plasma samples were stimulated. A study involving plasma samples from ten patients with celiac disease and ten healthy controls showed significant differences in mRNA TNF-α expression between experiments in which THP1 cells were stimulated by DNase-treated...
|
guest ::
