Bi9020c Evolutionary and comparative plant cytogenetics - practice

Faculty of Science
Autumn 2009
Extent and Intensity
0/2. 2 credit(s) (fasci plus compl plus > 4). Recommended Type of Completion: k (colloquium). Other types of completion: z (credit).
Teacher(s)
prof. Mgr. Martin Lysák, Ph.D., DSc. (seminar tutor)
RNDr. Terezie Malík Mandáková, Ph.D. (seminar tutor)
Mgr. Petr Mokroš, Ph.D. (seminar tutor)
Guaranteed by
prof. RNDr. Jiří Fajkus, CSc.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: prof. Mgr. Martin Lysák, Ph.D., DSc.
Prerequisites
Basics of cytogenetics and molecular or cell biology.
Course Enrolment Limitations
The course is only offered to the students of the study fields the course is directly associated with.

The capacity limit for the course is 20 student(s).
Current registration and enrolment status: enrolled: 0/20, only registered: 0/20
fields of study / plans the course is directly associated with
there are 8 fields of study the course is directly associated with, display
Course objectives
The subject of this practical course are classical as well as modern methods of plant molecular cytogenetics. Each practical task has a short theoretical introduction. The most important techniques demonstrated include: fluorescence in situ hybridization (FISH), fibre FISH, genomic in situ hybridization (GISH), DNA labelling by modified nucleotides, comparative chromosome painting, meiotic chromosomes in cytogenetics, immunolokalization of modified histones, fluorescence microscopy. The practical course follows up the lecture course Bi9020 Evolutionary and comparative plant cytogenetics. At the end of the practical course, students should acquire knowledge of the basic techniques of modern molecular plant cytogenetics.
Syllabus
  • Day 1
  • - direct and indirect labelling of DNA probes and their control on a gel (amplification and labelling of the pAL centromeric tandem repeats from A. thaliana using PCR; labelling of rDNA repeat and chromosome-specific BAC clones of A. thaliana using nick translation; labelling of genomic DNA by nick translation)
  • - microscopic preparations from root meristems (hybrids Festuca x Lolium: mitotic chromosomes) and anthers (A. thaliana: meiotic chromosomes)
  • - analysis of the slides using a fluorescence microscope (selection of suitable preparations for Day 2)
  • Day 2
  • - fluorescence in situ hybridization (FISH, chromosome painting)/genomic in situ hybridization (GISH), part I (making microscopic slides for FISH, preparation of hybridization probes, hybridization)
  • - theoretical and practical priciples of fluorescence microscopy, manipulation with fluorescence microscope and the corresponding software
  • - study of meiosis (meiotic stages analyzed using prepared slides using fluorescnce microscope, digital photographic documentation)
  • Day 3
  • - FISH, chromosome painting/GISH, part II (detection indirectly labelled DNA probes)
  • - analysis of slides using fluorescence microscope, digital photographic documentation (localization of tandem repeats on chromosomes and in nuclei of A. thaliana, chromosome painting: localization of BAC clones of A. thaliana on pachytene chromosomes and in interphase nuclei – chromosome territories; evaluation of GISH results – demonstration of inter-genomic translocations).
  • - processing of photographic outputs
  • Day 4
  • - making extended DNA fibres (A. thaliana) and FISH on extended DNA fibres, part I (directly labelled BAC clones made on Day 1)
  • - analysis of the results using a fluorescence microscope
  • - processing of photographic data
  • Day 5
  • - FISH on extended DNA fibres, part II
  • - analysis of the results using a fluorescence microscope
  • - processing of the photographic data
  • - conclusions and summary
Literature
  • SCHWARZACHER, T. and HESLOP-HARRISON. Practical in situ hybridization. Bios, 2000. info
Teaching methods
short theoretical introduction, lab work
Assessment methods
preparations, lab protocols, written test
Language of instruction
Czech
Follow-Up Courses
Further comments (probably available only in Czech)
The course is taught annually.
The course is taught: in blocks.
Note related to how often the course is taught: blok 5 dni.
The course is also listed under the following terms Autumn 2010 - only for the accreditation, Autumn 2010, Autumn 2011, Autumn 2011 - acreditation.
  • Enrolment Statistics (Autumn 2009, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2009/Bi9020c