C9042 Analysis of chromatin structure - practical training

Faculty of Science
Spring 2023
Extent and Intensity
0/0/2. 2 credit(s) (fasci plus compl plus > 4). Type of Completion: z (credit).
Teacher(s)
prof. RNDr. Jiří Fajkus, CSc. (seminar tutor)
Ing. Tereza Přerovská, Ph.D. (seminar tutor)
Mgr. Petra Procházková Schrumpfová, Ph.D. (seminar tutor)
doc. Mgr. Miloslava Fojtová, CSc. (seminar tutor)
Guaranteed by
prof. RNDr. Jiří Fajkus, CSc.
National Centre for Biomolecular Research – Faculty of Science
Supplier department: National Centre for Biomolecular Research – Faculty of Science
Prerequisites
NOW ( C9041 Struct. euk. chromosomes )
Basic knowledge of molecular biology and biochemistry, parallel or previous subscription of the course Structure and function of eukaryotic chromosomes (Bi9041), knowledge of laboratory safety instructions
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
The capacity limit for the course is 12 student(s).
Current registration and enrolment status: enrolled: 2/12, only registered: 0/12, only registered with preference (fields directly associated with the programme): 0/12
fields of study / plans the course is directly associated with
Course objectives
In the end of the course, students will be practically able to apply knowledge gained in the lecture course Structure and function of eukaryotic chromosomes (Bi9041). The aim of the course is to provide students with practical experience with methods used in analysis of the structure of chromosomes and chromatin. Main objectives: Student will be able: to isolate cell nuclei
to learn how to isolate and analyse histones
to learn MNase digestion of chromatin and analyse nucleosome spacing
Learning outcomes
After passing the course, a student will gain a practical experience in methods used in analysis of chromosome and chromatin structure.Student will be:
- practically acquainted with methods of preparation of cell nuclei;
- practically acquainted with methods of extraction and electrophoresis of histones;
- practically acquainted with methods of chromatin digestion with MNase;
- able to modify a protocol according to a material chosen.
Syllabus
  • 1. Isolation of cell nuclei from animal and plant tissues 2. Digestion of nuclei with micrococcal nuclease 3. Extraction of nucleosomal DNA 3. Analysis of nucleosomal DNA by electrophoresis 4. Preparation of mononucleosomal DNA from agarose gel 5. Mapping of nucleosome position using restriction enzymes or primer extension 6. Computer modelling of nucleosome position 7. Comparison of experimental and computer-predicted results 8. Extraction of chromatin proteins 9. Analysis of chromatin proteins by SDS-PAGE
Literature
  • FAJKUS, Jiří, Andrew R. LEITCH, Michael CHESTER and Eva SÝKOROVÁ. Evolution, Composition and Interrelated Functions of Telomeres and Subtelomeres: Lessons from Plants. Online. In Origin and Evolution of Telomeres. USA: Landes Biosciences, 2008. p. 114-127. ISBN 978-1-58706-309-1. [citováno 2024-04-23] info
  • FAJKUS, Jiří and Edward N. TRIFONOV. Columnar Packing of Telomeric Nucleosomes. Online. Biochemical and Biophysical Research Communications. California: Academic Press, 2001, roč. 280, No 4, p. 961-963. ISSN 0006-291X. [citováno 2024-04-23] info
Teaching methods
Practical laboratory training - intensive block teaching
Assessment methods
Written protocol emphasizing problems met during the project solving and discussion of the results. Final group discussion of the course experience.
Language of instruction
Czech
Further comments (probably available only in Czech)
The course is taught annually.
The course is taught: in blocks.
The course is also listed under the following terms Spring 2013, Spring 2014, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2022, Spring 2024, Spring 2025.
  • Enrolment Statistics (Spring 2023, recent)
  • Permalink: https://is.muni.cz/course/sci/spring2023/C9042