Bi5599c Applied Biochemistry and Cell Biology Methods - practice

Faculty of Science
Autumn 2020
Extent and Intensity
0/2/0. 2 credit(s) (plus extra credits for completion). Type of Completion: z (credit).
Taught in person.
Teacher(s)
Mgr. Jiřina Medalová, Ph.D. (seminar tutor)
RNDr. Vendula Pospíchalová, PhD. (seminar tutor)
Mgr. Petra Černochová (seminar tutor)
Guaranteed by
prof. Mgr. Vítězslav Bryja, Ph.D.
Department of Experimental Biology - Biology Section - Faculty of Science
Contact Person: Mgr. Jiřina Medalová, Ph.D.
Supplier department: Department of Experimental Biology - Biology Section - Faculty of Science
Timetable
Mon 9:00–10:50 D36/215
Prerequisites
Bi3030 Animal physiology && NOW ( Bi5599 App. Biochem. and Cell Methods )
The subject follows and belongs to the logical unit of practical courses of Animal Physiology and Special Methods of Animal Physiology.
Course Enrolment Limitations
The course is only offered to the students of the study fields the course is directly associated with.
fields of study / plans the course is directly associated with
there are 12 fields of study the course is directly associated with, display
Course objectives
The aim of this practicals is to teach students to: perform both the classical and modern techniques used in basic and applied research (see Syllabus); use up-to-date trends and modern techniques used in basic research of molecules, specific cells and cell populations; evolve their laboratory skills; decide what is the most appropriate assay with newly obtained methodological background; work in well-equipped facilities; use obtained knowledge and skills in experiments designed for their bachelor and diploma thesis
Learning outcomes
At the end of the course students should be able to: perform both the classical and modern techniques used in basic and applied research (see Syllabus); use up-to-date trends and modern techniques used in basic research of molecules, specific cells and cell populations; evolve their laboratory skills; decide what is the most appropriate assay with newly obtained methodological background; work in well-equipped facilities; use obtained knowledge and skills in experiments designed for their bachelor and diploma thesis
Syllabus
  • 1. Cultivation and use of cell cultures a) Lab work safety, how to use flow box, cell splitting, seeding and treatment - modification of beta catenin activity b) Transfection of cells by PEI polycation - assessment of transfection efficiency, TopFlash method for assessment of beta catenin activity 2. Gene expression evaluation by PCR a) Isolation of RNA by affinity purification, assessment of concentration and purity and stability of isolated RNA b) Reverse transcription of mRNA to cDNA c) Quantitative real time PCR for expression of beta catenin target genes, confirmation of reaction specificity by melting curve, methods for evaluation of results, counting of solution concentrations, cell suspensions dilution etc. 3. Assessment of protein expression by Western blotting a) Preparation and quantification of SDS lysates b) Separation of protein fragments by SDS PAGE c) Wet blotting of proteins to PVDF membrane d) Immunodetection of proteins - beta catenin and receptors involved to its regulation
Literature
  • Cell culture and upstream processing. Edited by M. Butler. 1st ed. New York: Taylor & Francis, 2007. xiv, 187. ISBN 9780415399692. info
  • Mammalian cell culture : essential techniques. Edited by Alan Doyle - J. B. Griffiths. Chichester: John Wiley & Sons, 1997. xv, 173. ISBN 0471970573. info
  • Cancer cell culture : methods and protocols. Edited by Simon P. Langdon. Totowa, N.J.: Humana Press, 2004. xii, 360. ISBN 1588290794. info
  • Animal cell culture :a practical approach. Edited by R. Ian Freshney. 2nd ed. Oxford: Oxford University Press, 1992. xix, 329 s. ISBN 0-19-963213-8. info
  • Cell biological appplications of confocal microscopy. Edited by Brian Matsumoto. 2nd ed. Amsterdam: Academic Press, 2002. xii, 507. ISBN 0125804458. info
  • Molecular cloning, A laboratory manual, Vol. 1-3, Eds. J. Sambrook and D. W. Russell, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York 2001
  • Short protocols in molecular biology, Vol. 1 and 2, 5th edition, John Wiley and Sons, Inc., 2002
  • Current protocols in cytometry, Vol. 1 and 2, Ed. J. P. Robinson, John Wiley and Sons, Inc., 1997
  • Methods in Cell Biology, Vol. 75, Cytometry, 4th edition, New developments, Eds. Z. Darzynkiewicz, M. Roederer, H. J. Tanke, Elsevier Academic Press, 2004
  • CROWTHER, John R. The ELISA guidebook. Totowa, N.J.: Humana Press, 2001. xi, 421. ISBN 0896037282. info
Teaching methods
According to the current measures of Masaryk University, the course will be taught as usually in three blocks. Each block will be introduces by short theoretic lecture, followed by practical experiments involving selected most important methods. Evaluation and conclusion of obtained results will be performed individually. If the hands-on teaching will be canceled, students will be prepared for their experiments via commented videos shared via MS Teams.
Assessment methods
In case of laboratory practical course must students complete given protocols with filled results and answers and pass the written test. In case of distant education instead of protocol must students write an assay on given issue and pass the test in IS.
Language of instruction
Czech
Follow-Up Courses
Further comments (probably available only in Czech)
Study Materials
The course is taught annually.
Information on course enrolment limitations: Na předmět se vztahuje povinnost registrace, bez registrace může být znemožněn zápis předmětu!
The course is also listed under the following terms Autumn 2018, Autumn 2019.
  • Enrolment Statistics (recent)
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