Bi7312 Practical course of molecular biology of eukaryotes

Faculty of Science
Autumn 2009
Extent and Intensity
0/2/0. 2 credit(s). Type of Completion: z (credit).
Teacher(s)
prof. RNDr. Jan Šmarda, CSc. (seminar tutor)
prof. Mgr. Petr Beneš, Ph.D. (seminar tutor)
doc. RNDr. Petr Vaňhara, Ph.D. (assistant)
Guaranteed by
prof. RNDr. Jan Šmarda, CSc.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: prof. RNDr. Jan Šmarda, CSc.
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, inoculation, 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 20 student(s).
Current registration and enrolment status: enrolled: 0/20, only registered: 0/20
fields of study / plans the course is directly associated with
Course objectives
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 split adherent and suspension cells, how to perform transient transfections of animal and/or human cells with exogenous genes/cDNAs using electroporation and calcium phosphate precipitation. In addition, they will learn how to document the presence and activity of exogenous gene(s) in transfected cells using various approaches targeting physical presence (immunoblotting, fluorescence microscopy) and activity (luciferase assay,beta-galactosidase assay) of their protein products.
Syllabus
  • 1. Splitting of suspension and adherent cells of BM2 and QT6 cell lines. 2. Transient transfection of BM2 cells with the plasmid cmvGFP by electroporation. Analysis of transfection efficiency by fluorescence microscopy. 3. Transient transfection of QT6 cells with the plasmids cmvbeta-gal, NdGE and Ew5luc by calcium precipitation follwed by these analyses: a) determination of the physical presence of Myb protein coded by the NdGE plasmid in transfected cells (SDS-PAGE, electroblotting, antibody detection b) determination of v-Myb protein activity in transfected cells (luciferase assay) c) determination of transfection efficiency by measurement of the beta-galactosidase activity in transfected cells 4. Study of cellular morphology in various phases of differentiation: Comparison of monoblasts and macrophages (cytocentrifugation, fixation, staining, light microscopy).
Literature
  • SAMBROOK, J., E.F. FRITSCH and T. MANIATIS. Molecular Cloning. A laboratory Manual. Second Edition. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, 1989. ISBN 0-87969-309-6. info
Teaching methods
- brief lectures followed by practical training - protocols describing the experiments and summarizing the results obtained are required
Assessment methods
Credits are given for active work at a bench, participation i each lesson and elaboration of detailed protocols describing the way of performing experiments and discussion of results and conclusions obtained.
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 taught: in blocks.
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 2010, Autumn 2011, Autumn 2011 - acreditation, Autumn 2012, Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020, autumn 2021, Autumn 2022, Autumn 2023, Autumn 2024.
  • Enrolment Statistics (Autumn 2009, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2009/Bi7312