1. komponenta

Lecture type	Total
Lectures	30
Practicum	45
Seminar	30

* Load is given in academic hour (1 academic hour = 45 minutes)

COURSE AIM:
The aim of the course is to teach students the fundamental principles and methods of modern molecular and cell biology research. Students will acquire knowledge of safe and proper laboratory practices, techniques for cell culture, analysis of genetic material manipulation, as well as the application of various methods for detection and quantification of nucleic acids and proteins. Special emphasis is placed on understanding technologies such as PCR, sequencing, hybridization techniques, polymorphism analysis, reporter genes, and epigenetic markers, along with their applications in forensics, anthropology, and biomedical research.
COURSE CONTENT:

LECTURES

1?2) Basics of laboratory work, categorization of laboratories and experimental organisms, laboratory safety rules, categorization into four biosafety levels depending on experimental models and transgenic organisms. Methods of sterilization, waste disposal, and personal protection.
3?4) Basics of cell culture work and reporter genes. Types of cultured cells, growth conditions, methods of foreign gene introduction, expression systems, vector structure, types of reporter genes, their applications and detection methods, flow cytometry.
5?6) DNA hybridization methods: basics of fluorescence in situ hybridization (FISH) and related techniques.
7?8) Analysis of polymorphisms in genomic and mtDNA: VNTR, STR, and SNP polymorphisms and their analysis in anthropology and forensics. Structure and analysis of hypervariable regions of mitochondrial DNA and their application. Paternity testing, population origin, and criminal investigations.
9?10) Sequencing: Sanger sequencing method, second-generation and third-generation sequencing. Advantages and disadvantages of each method and historical overview.
11?12) PCR and real-time PCR: method basics, types and design of primers, product visualization, and analysis of the result.
13?14) Protein analysis methods: protein isolation, methods for determining protein concentration, SDS-PAGE electrophoresis, Western blotting and detection of target proteins. Methods for studying protein interactions such as immunoprecipitation, FRET techniques, and the yeast two-hybrid method. Basics and applications of immunostaining.
15) Detection of epigenetic marks: methods for studying histone modifications and DNA methylation.

SEMINARS

1) Writing and presenting a research proposal.
2) Presentation of content and results from laboratory practice.

LABORATORY PRACTICE
1) Transfection of human cells with GFP vector ? the exercise includes working with cell culture, subculturing cells, calcium phosphate transfection method, analysis using fluorescence microscopy, image capture, and image processing in Photoshop.
2) Detection of the presence and potential aberrations of genes and/or chromosomes using the FISH method.
3) RT-qPCR for determining gene expression levels.
4 & 5) Protein analysis methods: Western blotting and immunostaining.
6) Application of methods ? practice at partner institutions.

1. Ambriović Ristov A. Metode u molekularnoj biologiji, Institut Ruđer Bošković 2007.
   Keith Wilson i John Walker: Principles and Techniques of Biochemistry and Molecular Biology, Cambridge University Press, 2011.
   Kathy Barker: At the bench: A laboratory navigator, Cold Spring Harbor Laboratory Press 2005.

Mandatory course - Regular study - Molecular Biology