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

Přírodovědecká fakulta
podzim 2022
Rozsah
0/24. 2 kr. (plus ukončení). Ukončení: z.
Vyučující
RNDr. Terezie Malík Mandáková, Ph.D. (cvičící)
Garance
prof. Mgr. Martin Lysák, Ph.D., DSc.
Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Kontaktní osoba: RNDr. Terezie Malík Mandáková, Ph.D.
Dodavatelské pracoviště: Národní centrum pro výzkum biomolekul – Přírodovědecká fakulta
Předpoklady
SOUHLAS
Prerequisites: basic knowledge in the field of cytogenetics and cytology (e.g., pipetting, preparation of solutions, microscopy).
Omezení zápisu do předmětu
Předmět je otevřen studentům libovolného oboru.
Předmět si smí zapsat nejvýše 10 stud.
Momentální stav registrace a zápisu: zapsáno: 0/10, pouze zareg.: 0/10, pouze zareg. s předností (mateřské obory): 0/10
Cíle předmětu
The participant will master basic techniques of plant cytogenetics (microscopic preps, basic fluorescence microscopy, chromosome painting, genomic in situ hybridization, image processing).
Výstupy z učení
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.
Osnova
  • 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
Literatura
    doporučená literatura
  • MANDÁKOVÁ, Terezie a Martin LYSÁK. Chromosome Preparation for Cytogenetic Analyses in Arabidopsis. Current protocols in plant biology. John Wiley & Sons, 2016, roč. 1, č. 1, s. 43-51. ISSN 2379-8068. Dostupné z: https://dx.doi.org/10.1002/cppb.20009. URL info
  • MANDÁKOVÁ, Terezie a Martin LYSÁK. Painting of Arabidopsis Chromosomes with Chromosome-Specific BAC Clones. Current protocols in plant biology. John Wiley & Sons, 2016, roč. 4, č. 2, s. 359-371. ISSN 2379-8068. Dostupné z: https://dx.doi.org/10.1002/cppb.20022. URL info
  • SCHWARZACHER, T. a HESLOP-HARRISON. Practical in situ hybridization. Bios, 2000. info
Výukové metody
Lectures followed by practical tasks in a cytogenetic laboratory.
Metody hodnocení
Colloquium. Successful completion is conditioned by a timely delivery of all required protocols and completion of the final interview.
Vyučovací jazyk
Angličtina
Navazující předměty
Další komentáře
Předmět je dovoleno ukončit i mimo zkouškové období.
Předmět je vyučován každoročně.
Výuka probíhá blokově.
Předmět je zařazen také v obdobích jaro 2012 - akreditace, podzim 2014, podzim 2015, podzim 2016, podzim 2017, podzim 2018, podzim 2019, podzim 2020, podzim 2021, podzim 2023.