1. General introduction: structure of nucleic acids, their importance and function, replication, transcription, translation. History of DNA diagnostics. 2. Amplification and the principle of polymerase chain reaction (PCR), individual steps of PCR, thermocyclers. PCR conditions: master mixes, template DNA, primers, buffers, dNTPs, types and kinds of polymerases. Methods of in vitro DNA propagation. 3. DNA cloning in practice, creation and importance of DNA libraries, DNA sequencing methods - classical and contemporary, blotting, NK databases, orientation and searching in NK databases. 4. Detection of amplified DNA in end-point PCR, agarose gels, polyacrylamide gels, electrophoresis, blotting, immunochromatography. 5. ELISA methods and their importance in diagnostics. 6. NK analysis by restriction analysis, application in diagnostics. Diagnostic methods derived from PCR, in situ PCR. 7. Real-time PCR detection of amplified DNA, principle of quantification, basic chemistry (Sybrgreen, TaqMan, etc.), molecular beacons, detection of point polymorphisms. 8. NK diagnostics in medicine: microbiology, virology, parasitology: practical application of NK diagnostics in direct detection of microorganisms. Detection of MTB, borrelia, chlamydia, HIV, viral hepatitis, herpes viruses, etc. 9. NK diagnostics in agriculture and forestry. 10. DNA diagnostics in food industry: diagnostics of genetically modified organisms in food and feed, species specification of meat products, cheese, European legislation on GMOs. 11. Diagnostics of NK in veterinary medicine. 12. Introduction of diagnostic methods, quality control (QC), quality assurance (QA), validation, certification, legislation. 13. Molecular biological diagnostic methods in forensic genetics.
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-
Alberts B, Johnson A, Lewis J, Morgan D, Raff M, Roberts K, Walter P. (2017). Molecular Biology of the Cell, 6th edition. Garland Science.
-
Drábek, J. a kol. (2012). Detekce nádorových biomarkerů v molekulárně-biologické laboratoři. Olomouc.
-
Drábek, J. a kol. (2022). Vybrané metody forenzní genetiky. Olomouc.
-
Hajdúch, M. (ed.) Drábek, J. (2022). Laboratory Techniques in Cellular and Molecular Medicine. Olomouc.
-
Slabý, O., et al. (2015). Molekulární medicína. Praha.
-
Šmarda, J., Doškař, J., Pantůček, R., Růžičková, V., & Koptíková, J. (2005). Metody molekulární biologie. Brno.
-
Vacek, J., Ostatná, V. (2021). Bioelektrochemie. Olomouc.
|