Bi7312 Practical course of molecular biology of eukaryotes

Faculty of Science
Autumn 2019
Extent and Intensity
0/2/0. 2 credit(s). Type of Completion: z (credit).
Teacher(s)
doc. Mgr. Petr Beneš, Ph.D. (seminar tutor)
Mgr. Lucia Knopfová, Ph.D. (seminar tutor)
Mgr. Jarmila Navrátilová, Ph.D. (seminar tutor)
RNDr. Martin Šrámek, Ph.D. (assistant)
Mgr. Monika Dúcka, Ph.D. (assistant)
Ing. Barbora Adamová, Ph.D. (assistant)
Mgr. Kamila Říhová, Ph.D. (assistant)
Mgr. Ivana Kupčíková, DiS. (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.
Bi7312/02: No timetable has been entered into IS.
Bi7312/03: No timetable has been entered into IS.
Bi7312/04: No timetable has been entered into IS.
Bi7312/05: No timetable has been entered into IS.
Bi7312/06: No timetable has been entered into IS.
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: 0/30, only registered: 0/30
fields of study / plans the course is directly associated with
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.
The course is also listed under the following terms Autumn 2007 - for the purpose of the accreditation, Autumn 2010 - only for the accreditation, Autumn 2002, Autumn 2003, Autumn 2004, Autumn 2005, Autumn 2006, Autumn 2007, Autumn 2008, Autumn 2009, Autumn 2010, Autumn 2011, Autumn 2011 - acreditation, Autumn 2012, Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2020, autumn 2021, Autumn 2022, Autumn 2023, Autumn 2024.
  • Enrolment Statistics (Autumn 2019, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2019/Bi7312