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Lecturer(s)
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Course content
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Lectures: 1: Introduction. Nucleic acid isolation requirements, source material, principles of techniques. Analysis and quantification of isolated nucleic acids 2: Electrophoresis principles, types of gels, electrophoretic buffers, factors influencing electrophoresis. Detection of separated biomacromolecules. Modifications of electrophoretic techniques (PFGE, SSCP, TGGE, DGGE et al.) 3: Enzymes in molecular biology, enzymatic manipulation of nucleic acids, in vitro enzyme modifications. Polymerases, restriction endonucleases. Restriction fragment length polymorphism (RFLP). Application of RFLP in food hygiene 4: Polymerase chain reaction (PCR) - principles, conditions, components, basic principles of primer design, application of PCR in food hygiene 5: Variants of PCR (RT-PCR, nested PCR, multiplex PCR et al.). Real-time PCR (qPCR). LAMP. DNA sequencing. Principles, components, application. Automatic sequencer. Basic principles of sequencing data evaluation 6: Next-generation sequencing (NGS), principles of NGS. Whole-genome sequencing. NGS in food hygiene 7: Genetically modified organisms (GMO). Principles and examples of genetical modifications, GMO in food hygiene Practical lessons: 1. DNA isolation from fillets 2. Quantification of extracted DNA 3. Polymerase chain reaction (PCR) I - primer design 4. Polymerase chain reaction (PCR) II - setup 5. Polymerase chain reaction (PCR) III - execution 6. Electrophoretic separation of PCR products I - setup 7. Electrophoretic separation of PCR products II - execution 8. Purification of PCR products I - setup 9. Purification of PCR products II - execution 10. Processing of sequencing data, MEGA software 11. GenBank and BOLD databases 12. Evaluation of sequencing data 13. Conclusions and assessment of results 14. Awarding of credit
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Learning activities and teaching methods
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Lectures, Laboratory and desk-based work
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Learning outcomes
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The aim of the course is to introduce students to selected basic methods of molecular biology, their principles, and modifications. Furthermore, students obtain information about basic problems in food hygiene solvable by molecular biology methods and are introduced to selected bioinformatics tools and databases. During practicals, students solve a practical problem related to food hygiene (identification of a fish meat sample utilizing DNA analysis).
1) Theoretical knowledge The student is able to: - explain principles of basic methods of molecular biology, - describe the execution of basic methods of molecular biology and their practical application, - describe problems in food hygiene solvable by methods of molecular biology,. - understand veterinary public health issues, e.g. epidemiology, transboundary epizootic diseases, zoonotic and food-borne diseases, emerging and re-emerging diseases, food hygiene and technology. 2) Practical skills The student is able to: - process samples intended to be analysed by methods of molecular biology according to the principles of proper laboratory practice, - utilize basic bioinformatics tools and databases necessary to work with data sequencing, - perform the isolation of DNA from a sample properly and to perform PCR and gel electrophoresis, - based on sequence data analysis, identify the taxonomic origin of the analysed foodstuff correctly. 3) Competences The student is able to: - work effectively as a member of a multi-disciplinary team in the delivery of services - demonstrate an ability of lifelong learning and a commitment to learning and professional development. This includes recording and reflecting on professional experience and taking measures to improve performance and competence, - take part in self-audit and peer-group review processes in order to improve performance, - collect, preserve and transport samples, select appropriate diagnostic t ests, interpret and understand the limitations of the test results, - perform inspection of food and feed including post-mortem inspection of food producing animals and inspection in the field of related food technology, - choose on their own a suitable method for solving a given food hygiene problem, including assessment of possibilities and limits of the respective methods, - interpret results of molecular-biological analysis of food sample.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Oral exam, Written credit test,
Credit requirements: - attendance at all practicals - preparation of assignment protocols - correct identification of a sample based on analysis of sequence data Exam requirements: - to be admitted to the exam, each student must obtain the credit first - acceptable knowledge level in two fields (molecular biology in food hygiene + isolation of nucleic acids + PCR; electrophoresis + enzyme methods + DNA sequencing + GMO) - the student picks two questions randomly (one from each field), has 30 minutes available to prepare
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Recommended literature
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Aly Farag El Sheikha, Robert Levin, Jianping Xu. Molecular techniques in food biology: safety, biotechnology, authenticity and traceability. Hoboken, NJ, 2018. ISBN 978-1-11-937459-6.
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Šmarda J. a kol. Metody molekulární biologie. ISBN 80-210-3841-1.
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