S2003 Methods in plant cytogenetics and cytogenomics II. - practical course

Faculty of Science
Spring 2021
Extent and Intensity
0/24. 2 credit(s) (plus extra credits for completion). Type of Completion: z (credit).
Teacher(s)
RNDr. Terezie Mandáková, Ph.D. (lecturer)
Guaranteed by
prof. Mgr. Martin Lysák, Ph.D., DSc.
National Centre for Biomolecular Research - Faculty of Science
Contact Person: RNDr. Terezie Mandáková, Ph.D.
Supplier department: National Centre for Biomolecular Research - Faculty of Science
Prerequisites
S2002 Methods plant cytogenetics 1
The completion of the basic course "Methods in plant cytogenetics and cytogenomics I. (S2002)" is a prerequisite for enrolling this advanced course.
Course Enrolment Limitations
The course is offered to students of any study field.
The capacity limit for the course is 10 student(s).
Current registration and enrolment status: enrolled: 0/10, only registered: 0/10, only registered with preference (fields directly associated with the programme): 0/10
Course objectives
Students will get acquainted with preparation of extended DNA fibres (fibre FISH), immuno localization of modified histones, immuno-labeling of 5-methyl cytosine, and FISH to immuno preparations. Advanced methods of fluorescence microscopy and digital photomicrography will be practised.
Learning outcomes
Student will be able to (i) prepare extended DNA fibres, (ii) prepare DNA probes, (iii) carry out fibre FISH, immuno-localization of modified histones, immuno-labeling of 5-methyl cytosine and FISH to immuno preparations, (iv) analyze preparations using fluorescence microscopy, (v) photograph chromosomes and cell nuclei, and finally (vi) analyze and process the acquired photographs.
Syllabus
  • Annotation This advanced course follows Methods in plant cytogenetics I. Aim of the course is to make students familiar with the principles and applications of DNA-DNA fluorescence in situ hybridization (FISH) and immuno-localisation as tools for studying the chromatin and nuclear organization of higher plants. Students will get acquainted with preparation of extended DNA fibres (fibre FISH), immuno-localization of modified histones, immuno-labeling of 5-methyl cytosine, and FISH to immuno-preparations. Advanced methods of fluorescence microscopy and digital photomicrography will be practised. Syllabus Day 1 - introduction in theoretical and practical principles of nuclei isolation and DNA fibre FISH - nuclei isolation from leaves - nuclei extension and fibre FISH hybridization with probes corresponding to ribosomal DNA repeats Day 2 - introduction in multilayer detection of the fluorescence signals and immuno-localisation of modified histones and methylated DNA - fibre FISH multilayer detection - immuno-localisation of modified histones H3metK4 and H3metK9: PART I - immuno-labelling of 5-methyl cytosine on mitotic and meiotic chromosomes - fluorescence microscopy of fibre FISH and immuno-labelled methylcytosine Day 3 - immuno-localisation of modified histones H3metK4 and H3metK9: PART II - fluorescence microscopy of modified histones - advanced analysis using fluorescence microscopy, image capture - computer processing of microscope images (Adobe Photoshop) - conclusions
Literature
    recommended literature
  • SCHWARZACHER, T. and HESLOP-HARRISON. Practical in situ hybridization. Bios, 2000. info
Teaching methods
Lectures followed by practical tasks in a cytogenetic laboratory.
Assessment methods
Colloquium. Successful completion is conditioned by a timely delivery of all required protocols and completion of the final interview.
Language of instruction
English
Further Comments
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 spring 2012 - acreditation, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2022.
  • Enrolment Statistics (recent)
  • Permalink: https://is.muni.cz/course/sci/spring2021/S2003