LEARNING OUTCOMES
Aim of the course:
The theoretical lectures and the laboratory exercises of the course introduce students to the basic concepts, principles and methods of molecular biology used in laboratories providing services to the public and in research protocols. The lectures and the laboratory exercises will operate simultaneously as much as possible and the monitoring of students’ knowledge and progress will be provided by multiple choice tests and feedback during the laboratory exercises. Students upon the end of the course should have obtained the knowledge and expertise to attend special background courses and laboratory exercises in the following semesters, as well as to know and understand both the basic concepts of molecular biology and the theoretical and practical background of the basic methods of molecular biology. Students after the end of the semester, must have acquired good theoretical and practical knowledge of modern methods.
SYLLABUS
Lectures:
1. Structure and properties of DNA and RNA: Central dogma of molecular biology -DNA- Structure and function. Chromatin Forms-Chromosomes. DNA denaturation, rearrangement and DNA ligation. Nucleic acid hybridization. Supercoiled DNA, topoisomerases. Bacterial and eukaryotic chromosomes. Nucleosomes, active chromatin. Relationship between genome size and organ complexity. Repetitive DNA. The genetic material of viruses and bacteriophages.
2. Recombinant DNA technology: Restriction enzymes (endonucleases). Artificial ligation of DNA fragments. DNA mapping using restriction enzymes.
3. Cloning, cloning vectors.
4. Nucleic acid hybridization: Detection and characterization of specific DNA sequences, probes, hybridizations using Southern, Northern, etc.
5. Determination methods of primary DNA structure. Research programs for determining whole genomes of organisms.
6. Polymerase chain reaction (PCR) and its applications. Real time polymerase chain reaction (real time PCR) and its applications
7. DNA replication: Protein factors and replication enzymes. Mechanisms of DNA replication in prokaryotic and eukaryotic organisms. The replication mechanisms in bacteriophage DNA as well as in DNA and RNA viruses.
8. Transcription: Transcription enzymes. Promoter sequences. Transcription mechanism in prokaryotic organisms. RNA maturation. Regulatory mechanisms.
9. Transcription in eukaryotic organisms: Specific features. Transcription factors. RNA maturation. Regulation mechanisms of transcription.
10. Protein biosynthesis (translation): Genetic code, instability, repression, mRNA origin and evolution of the genetic code. Mechanisms of protein synthesis. Structure and functionality of tRNAs. Structure and functionality of ribosomes. Role of ribosomal RNA in protein synthesis.
11. Gene expression study using molecular biology methods such as RT-PCR and qRT-PCR as well as microarrays and RNA-sequencing technology
12. Transgenic animals and genetic modification
13. Bioinformatics in molecular biology
Laboratory exercises:
14. Genomic DNA isolation of from eukaryotic cells
15. Restrictive enzymes (endonucleases) – digestion and genomic DNA electrophoresis
16. Plasmid isolation from bacterial cells
17. Restriction enzyme maps-Electrophoresis of plasmid digestion products
18. RNA isolation
19. Nucleic acid electrophoresis (DNA, RNA)
20. Southern, Northern, in situ hybridization (ISH), Fluorescent in situ hybridization (FISH)
21. Polymerase chain reaction (PCR)
22. Real time PCR reaction
23. Reverse transcriptase reaction, RT-PCR, qRT-PCR
24. Hybridization Microarrays, RNA Sequencing
25. Introduction of recombinant plasmid into bacterial cells and selection of clones with antibiotic in solid medium cultures
26. Sanger sequencing, NGS (Next Generation sequencing).