Molecular Biology and Gene Technology I

Data

Official data in SubjectManager for the following academic year: 2024-2025

Topic

Course lectures present the theoretic background and practical utilization of current and future-proof methodology of molecular biology, and within that genetic engineering. The course includes the following methodologies: various PCR types, restriction endonuclease digestion, molecular cloning, RNA interference and CRISPR/Cas9, transduction and transfection of pro- and eukaryotic cells, expression systems, development of genetically modified cells and organisms, gene therapy applications. During course seminars the students will familiarize with basic molecular biology and within that genetic engineering-related trouble-shooting. During course practices pipetting and precision skills, sterile techniques, master-mix preparation, serial dilutions etc. will be practiced. Solving simple practical issues will help to gain deep understanding and practical knowledge. Having fulfilled this molecular biology and genetic engineering-based course the graduates will get hands-on- expertise that will aid their efforts on the labor-market for both academic and industrial positions.

Lectures

• 1. DNA, gene structure - Kvell Krisztián
• 2. Transcription, translation, central dogma - Kvell Krisztián
• 3. Genome organisation (prokaryotes, eukaryotes) - Kvell Krisztián
• 4. RNA species focusing on mRNA and miRNA - Kvell Krisztián
• 5. Extracellular vesicle subtypes - Kvell Krisztián
• 6. Extracellular vesicles as carriers - Kvell Krisztián
• 7. Lipid nanoparticles as carriers - Kvell Krisztián
• 8. Enzyme tools: restriction endonucleases - Kvell Krisztián
• 9. Enzyme tools: ligase, topoisomerase, klenow - Kvell Krisztián
• 10. Enzyme tools: reverse transcriptase - Kvell Krisztián
• 11. Enzyme tools: heatstable polymerases - Kvell Krisztián
• 12. PCR: conventional - Kvell Krisztián
• 13. PCR: quantitative - Kvell Krisztián
• 14. PCR: absolute - Kvell Krisztián
• 15. Consultation - Kvell Krisztián
• 16. Midterm evaluation - Kvell Krisztián
• 17. Amplification-based high-throughput assay (Quantstudio) - Kvell Krisztián
• 18. Amplification-free high-throughput assay (Nanostring) - Kvell Krisztián
• 19. Plasmids in nature - Kvell Krisztián
• 20. Plasmids in the lab - Kvell Krisztián
• 21. Phages in nature - Kvell Krisztián
• 22. Phages in the lab - Kvell Krisztián
• 23. Bacterial transformation: calcium-phosphate - Kvell Krisztián
• 24. Bacterial transformation: electroporation - Kvell Krisztián
• 25. Bacterial antibiotic resistance in nature - Kvell Krisztián
• 26. Bacterial antibiotic resistance in the lab - Kvell Krisztián
• 27. Consultation - Kvell Krisztián
• 28. Final evaluation - Kvell Krisztián

Practices

• 7. Lipid nanoparticle production with Lipofectin I - Kvell Krisztián
• 6. Extracellular vesicle analysis with Nanosight - Kvell Krisztián
• 5. Extracellular vesicle enrichment - Kvell Krisztián
• 4. RNA isolation with silica membrane II - Kvell Krisztián
• 3. RNA isolation with silica membrane I - Kvell Krisztián
• 2. DNA isolation with silica membrane II - Kvell Krisztián
• 1. DNA isolation with silica membrane I - Kvell Krisztián
• 8. Lipid nanoparticle production with Lipofectin II - Kvell Krisztián
• 9. Reverse transcription using RT enzyme I - Kvell Krisztián
• 10. Reverse transcription using RT enzyme II - Kvell Krisztián
• 11. Preparing conventional PCR - Kvell Krisztián
• 12. Evaluating conventional PCR - Kvell Krisztián
• 13. Preparing quantitative PCR - Kvell Krisztián
• 14. Evaluating quantitative PCR - Kvell Krisztián
• 15. Consultation - Kvell Krisztián
• 16. Consultation - Kvell Krisztián
• 17. Evaluating Quantstudio results - Kvell Krisztián
• 18. Evaluating Nanostring results - Kvell Krisztián
• 19. Starting bacterial cultures with antibiotics - Kvell Krisztián
• 20. Evaluating bacterial cultures with antibiotics - Kvell Krisztián
• 21. Starting bacterial cultures with phages - Kvell Krisztián
• 22. Harvesting bacterial cultures with phages - Kvell Krisztián
• 23. Heat-shock transformation of bacterial cultures - Kvell Krisztián
• 24. Plating transformed bacteria, colony evaluation - Kvell Krisztián
• 25. Plasmid isolation with silica membrane I - Kvell Krisztián
• 26. Plasmid isolation with silica membrane II - Kvell Krisztián
• 27. Consultation - Kvell Krisztián
• 28. Consultation - Kvell Krisztián

Seminars

• 4. Concept of lipid nanoparticle production - Kvell Krisztián
• 3. Concept of extracellular vesicle analysis - Kvell Krisztián
• 2. Concept of kit-based RNA isolation - Kvell Krisztián
• 1. Concept of kit-based DNA isolation - Kvell Krisztián
• 5. Planning the use of enzymes in the lab - Kvell Krisztián
• 6. Planning a conventional PCR - Kvell Krisztián
• 7. Planning a quantitative PCR - Kvell Krisztián
• 8. Consultation - Kvell Krisztián
• 9. Planning the use of high-throughput assays - Kvell Krisztián
• 10. Planning an antibiogram - Kvell Krisztián
• 11. Planning bacterial phage transformation - Kvell Krisztián
• 12. Planning bacterial plasmid transformation - Kvell Krisztián
• 13. Concept of plasmid isolation - Kvell Krisztián
• 14. Consultation - Kvell Krisztián

Reading material

Obligatory literature

Cox, Doudna, O’Donnel: Molecular Biology: Principles and Practice, Freeman , 2nd Edition, 2015, ISBN:1464126143

Mitra, Sandhya: Genetic Engineering: Principles and Practice, MC GRAW HILL INDIA, 2015, ISBN-10 : 9789339203535, ISBN-13 : 978-9339203535

Literature developed by the Department

Notes

Recommended literature

Krebs, Goldstein, Kilpatrick: Lewin’s Essential Genes, Jones and Bartlett, 4th Edition, 2020, ISBN:1284173135

Conditions for acceptance of the semester

Labnotes submitted by end of semester

Mid-term exams

Single choice questions prepared according to the subject of the lectures. The written exam test might contain minimum requirement questions where the wrong answers could mean failure on the exam.

Making up for missed classes

Not possible.

Exam topics/questions

https://gytk.pte.hu/en/menupont/10537

Examiners

• Kvell Krisztián

Instructor / tutor of practices and seminars

• Hairui Ou
• Nemes Balázs