1. SylabUZ
2. Faculty of Exact and Natural Sciences
3. winter term 2019/2020
4. WNB - oferta ERASMUS - Erasmus programme
5. Biological Molecules Labeling Technique

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Biological Molecules Labeling Technique - course description

Aim of the course

The aim of education is to acquire by the student the theoretical knowledge about the application of modern biological molecules labeling techniques as well as basic practical skills in the techniques used in the biological molecule, labeling with particular emphasis on the documentation techniques. Students have the opportunity to work with the group and organize common tasks as well take responsibility for the equipment.

Prerequisites

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The basic knowledge of biochemistry, cell biology and genetics.

Scope

Lecture: Isotopic and non-isotopic methods for labeling of the nucleic acids, proteins, lipids, carbohydrates and other biologically active substances. Non-isotopic techniques for the fractionation and labelling of biological macromolecules. The labeling in vitro, in vivo and in silico. Cell organelle markers. Methods of signal detection after labeling of biological molecules. Electrophoretic and immunocytochemical techniques. The nucleic acids hybridization techniques. Methods: Southern blot, Northern blot, DNA microarrays and hybridization in situ. Plasmids labeling (nick translation, random priming). End-labeling of DNA. Restriction maps. Methods for quantitative determination of the labeled molecules. Immunochemical methods. Radioimmunological methods (RIA). Isotopes: 32P, 33P, 35S as well as 14C and 3H. Fluorescent methods (FIA). Fluorophores. Flow cytometry, FACS, FISH techniques, DNA and RNA sequencing, peptide and tissue microarrays. Immunoenzymatic methods (EIA). Western blot. ELISA test. Chemiluminescent methods. Quantity and quality practical analysis of the proteins in cell lysates by Western blot and chemiluminescence. Analysis of the location of proteins in human cells by fluorophore labeling and visualization in a confocal microscope.

Laboratory: Western blot analysis: Preparation of needed buffers and solutions for WB; Protein preparation - Cell lysis; SDS PAGE electrophoresis; Transferring proteins to a membrane; Blocking nonspecific sites; Primary and secondary antibodies; Chemiluminescent detection; Densitometry analysis. Application of confocal microscopy.

DNA sequencing (Sanger method); dyes used for labeling of biomolecules;  MACS Cell Separation (Miltenyi Biotec); Densitometry analysis (Image Lab Software, Image J Software).

Teaching methods

Lecture: giving method - lecture in the form of a multimedia presentation in the classroom;

Laboratory: giving method: discussion on the application of analytical methods; practical method: lab exercises with the use of selected: biological molecules labeling techniques, methods for documenting the results and bioinformatics programs.

Learning outcomes and methods of theirs verification

Outcome description	Outcome symbols	Methods of verification	The class form

Assignment conditions

Lecture: provided credit is to get positive results from essey. Least 50% of scored points are required to get the pass mark credit.

Laboratory: provided credit is class attendance and getting positive results of the test, and credit of all written reports from performed laboratory experiments. The final mark consists of the average sum of all of the passed partial marks.

Recommended reading

1.       J.M. Berg, J.L.Tymoczko, L. Stryer, Biochemistry, Seventh Edition Hardcover 2010

2.       D.L. Nelson, M.M. Cox Lehninger Principles of Biochemistry Hardcover, 2012

3.       T.A. Brown Genomes 3 2006

4.       B. Alberts, D. Bray, K. Hopkin, A. Johnson, J Lewis, M. Raff, K. Roberts, P. Walter Molecular Biology of the Cell Hardcover 2014

Further reading

1.       Selected current research and review articles.

Notes

Modified by dr hab. inż. Dżamila Bogusławska, prof. UZ (last modification: 08-05-2019 15:58)