PřF:Bi7312 Eukaryotic cell in practice - Course Information

Bi7312 Practical course of molecular biology of eukaryotes

Extent and Intensity

0/2/0. 2 credit(s). Type of Completion: z (credit).
In-person direct teaching

Teacher(s)

doc. Mgr. Petr Beneš, Ph.D. (seminar tutor)
Mgr. Lucia Knopfová, Ph.D. (seminar tutor)
Mgr. Jarmila Navrátilová, Ph.D. (seminar tutor)
Mgr. Jiří Kohoutek, Ph.D. (seminar tutor)
Mgr. Andrea Rousová (assistant)
Mgr. Jana Pokludová (assistant)
Mgr. Matěj Přikryl (assistant)

Guaranteed by

doc. Mgr. Petr Beneš, Ph.D.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: doc. Mgr. Petr Beneš, Ph.D.
Supplier department: Department of Experimental Biology – Biology Section – Faculty of Science

Timetable of Seminar Groups

Bi7312/01: No timetable has been entered into IS. L. Knopfová
Bi7312/02: No timetable has been entered into IS. J. Navrátilová, M. Přikryl
Bi7312/03: No timetable has been entered into IS. J. Pokludová, A. Rousová
Bi7312/04: No timetable has been entered into IS. L. Knopfová
Bi7312/05: No timetable has been entered into IS. J. Kohoutek
Bi7312/06: No timetable has been entered into IS. P. Beneš

Prerequisites

NOW( Bi7090 Eukaryotic cells )
The course requires basic practical skills of microbiology, biochemistry and molecular biology as, for example, preparation of buffers, cultivation media, sterile liquide handling, manipulation with automatic pipettes, centrifugation and so on.

Course Enrolment Limitations

The course is only offered to the students of the study fields the course is directly associated with.

The capacity limit for the course is 30 student(s).
Current registration and enrolment status: enrolled: 29/30, only registered: 0/30

fields of study / plans the course is directly associated with

• Bioanalytical Laboratory Diagnostics in Medicine - Medical Genetics and Molecular Diagnostics (programme PřF, N-LGM)
• Human Biology (programme PřF, N-BCL)
• Medical Genetics and Molecular Diagnostics (programme PřF, N-BI)
• Molecular Biology and Genetics (programme PřF, N-EXB)
• Molecular Biology and Genetics (programme PřF, N-MBG)

Course objectives

The aim of this practical course it to make students familiar with basic methodology of analysis of cell proliferation, differentiation, migration and cell death in model cancer cell lines.

Learning outcomes

At the end of the course students acquire practical skills with basic techniques allowing manipulations with animal and human cells in vitro. They will understand how to perform genetic modification of cells in culture, how to analyze cell morphology, proliferation, differentiation, migration and apoptosis.

Syllabus

• 1. Splitting of suspension and adherent mammalian cells, 2D and 3D cultivation. 2. Transient transfection of mammalian cells with the plasmid cmvGFP by electroporation. Analysis of transfection efficiency by fluorescence microscopy and flow-cytometry. 3. Analysis of activity of transcription factors (transient transfection, assays of beta-galactosidase and luciferase activities). 4. Study of cellular morphology, protein expression, protein localisation in various phases of cellular differentiation (cytocentrifugation, cell fixation and staining, light microscopy, fluorescence microscopy, cell fractionation, protein electrophoresis and immunoblotting, imunocytofluorescence, phagocytosis). 5. Analysis of cellular markers of apoptosis (staining of viable cells by eosin dye exclusion, analysis of nuclear morphology, PARP cleavage, caspase 3 activation, phosphatidylserine externalization, mitochondrial membrane potential evaluation, analysis of reactive oxygen species by flow-cytometry). 6. Determination of cell proliferation, migration and senescence (cell counting, crystal violet staining, cell cycle analysis, real-time cell proliferation and migration monitoring, SA-beta-gal staining). 7. Determination of cell viability in 3D models (spheroids) using confocal microscopy of calcein/propidium iodide-stained spheroids, MTT and ATP assays. Immunohistochemistry.

Literature

• GREEN, Michael R. and Joseph SAMBROOK. Molecular cloning : a laboratory manual. 4th ed. New York [N.Y.]: Cold Spring Harbor Laboratory Press, 2012, xxxiii, s. ISBN 9781936113422. info
• FRESHNEY, R. Ian. Culture of animal cells : a manual of basic technique. 5th ed. Hoboken, N.J.: Wiley-Liss, 2005, xxvi, 642. ISBN 0471453293. info
• Apoptosis methods and protocols. Edited by Hugh J. M. Brady. Totowa, N.J.: Humana Press, 2004, xi, 304. ISBN 0896038734. info

Teaching methods

brief lectures followed by practical training, preparation of final group project

Assessment methods

Credits are given for active work at a bench, participation i each lesson, preparation of final group project.

Language of instruction

Czech

Further Comments

Study Materials
The course can also be completed outside the examination period.
The course is taught annually.

• Enrolment Statistics (recent)
  
• Permalink: https://is.muni.cz/course/sci/autumn2024/Bi7312