S2002 Methods in plant cytogenetics and cytogenomics I. - practical course

Faculty of Science
Autumn 2023
Extent and Intensity
0/24/0. 2 credit(s) (plus extra credits for completion). Type of Completion: z (credit).
Teacher(s)
RNDr. Terezie Malík Mandáková, Ph.D. (seminar tutor)
Guaranteed by
RNDr. Terezie Malík Mandáková, Ph.D.
National Centre for Biomolecular Research – Faculty of Science
Contact Person: RNDr. Terezie Malík Mandáková, Ph.D.
Supplier department: National Centre for Biomolecular Research – Faculty of Science
Prerequisites
SOUHLAS
Prerequisites: basic knowledge in the field of cytogenetics and cytology (e.g., pipetting, preparation of solutions, microscopy).
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: 5/10, only registered: 0/10, only registered with preference (fields directly associated with the programme): 0/10
Course objectives
The participant will master basic techniques of plant cytogenetics (microscopic preps, basic fluorescence microscopy, chromosome painting, genomic in situ hybridization, image processing).
Learning outcomes
Student will be able to (i) prepare microscopic preparations containing mitotic and meiotic chromosomes, (ii) label isolated DNA by haptens and fluorochromes, (iii) prepare DNA probes, (iv) carry out fluorescence and genomic in situ hybridization (FISH, GISH) of the labeled DNA probes to chromosomes and cell nuclei, (v) analyze preparations using fluorescence microscopy, (vi) photograph chromosomes and cell nuclei, and finally (vii) analyze and process the acquired photographs.
Syllabus
  • Annotation This practical course will demonstrate classical as well as state-of-the-art cytogenetic techniques. The accent is put on cytogenetic methods beneficial to taxonomic, biosystematic and phylogeographic studies. Students will get acquainted with preparation of mitotic and meiotic chromosomes, chromosome counting, the course of male meiosis as well as with principles of light and fluorescence microscopy. Molecular cytogenetic techniques will include DNA probe labelling, fluorescence in situ hybridization (FISH), genomic in situ hybridization (GISH) and chromosome painting. Digital processing of captured chromosome images will be demonstrated. Syllabus Day 1 - introduction - preparation of mitotic chromosomes from root tip meristems (Nicotiana, Festuca x Lolium hybrids) and preparation of meiotic chromosome spreads from anthers - theoretical and practical principles of light and fluorescence microscopy - analysis of chromosome preparations using fluorescence microscopy (selection of suitable slides for Day 2) - direct and indirect labelling of DNA probes (amplification of the Arabidopsis-type telomere repeat by PCR; labelling of ribosomal DNA repeats, chromosome-specific BAC clones, and genomic DNA using nick translation) Day 2 - introduction - fluorescence in situ hybridization (FISH), chromosome painting, genomic in situ hybridization (GISH): PART I (slide pre-treatment, preparation of hybridization mix, probe denaturation) - stages of mitosis and meiosis (movie, meiosis analysed using phase and fluorescence microscopy, digital microscope photography) Day 3 - introduction - FISH, chromosome painting, GISH: PART II (fluorescence detection of hybridized DNA probes) - fluorescence microscopy, image capture - computer processing of microscope images (Adobe Photoshop) - conclusions
Literature
    recommended literature
  • MANDÁKOVÁ, Terezie and Martin LYSÁK. Chromosome Preparation for Cytogenetic Analyses in Arabidopsis. Current protocols in plant biology. John Wiley & Sons, 2016, vol. 1, No 1, p. 43-51. ISSN 2379-8068. Available from: https://dx.doi.org/10.1002/cppb.20009. URL info
  • MANDÁKOVÁ, Terezie and Martin LYSÁK. Painting of Arabidopsis Chromosomes with Chromosome-Specific BAC Clones. Current protocols in plant biology. John Wiley & Sons, 2016, vol. 4, No 2, p. 359-371. ISSN 2379-8068. Available from: https://dx.doi.org/10.1002/cppb.20022. URL info
  • 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
Follow-Up Courses
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, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020, autumn 2021, Autumn 2022.
  • Enrolment Statistics (recent)
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