C7235 Fluorescence methods in life sciences - excercise

Faculty of Science
autumn 2021
Extent and Intensity
0/2/0. 2 credit(s) (plus extra credits for completion). Recommended Type of Completion: k (colloquium). Other types of completion: z (credit).
Teacher(s)
doc. Mgr. Ctirad Hofr, Ph.D. (seminar tutor)
Ing. Tomáš Janovič, Ph.D. (seminar tutor)
Mgr. Pavel Veverka, Ph.D. (seminar tutor)
Mgr. Martin Stojaspal, Ph.D. (seminar tutor)
Mgr. Tomáš Brom (seminar tutor)
Guaranteed by
prof. RNDr. Jiří Fajkus, CSc.
National Centre for Biomolecular Research – Faculty of Science
Contact Person: doc. Mgr. Ctirad Hofr, Ph.D.
Supplier department: National Centre for Biomolecular Research – Faculty of Science
Prerequisites (in Czech)
NOW( C7230 Fluorescence methods )
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 30 student(s).
Current registration and enrolment status: enrolled: 2/30, only registered: 0/30, only registered with preference (fields directly associated with the programme): 0/30
fields of study / plans the course is directly associated with
there are 7 fields of study the course is directly associated with, display
Course objectives
The main objective of the practical training course is to acquire practical experience with modern spectroscopic methods employed in the analysis of protein-protein and protein-DNA interactions. From the methodical approaches, the main emphasis is put on highly sensitive fluorescent methods and their practical use in biological laboratories. The practical one week course, after finishing lectures, enables trainees to corroborate their theoretical knowledge about methodical principles using modern instrumentation of research laboratories. Training is focused on the effective development of skills required for application of specialized techniques. These techniques have become indispensable for recent tasks of basic and applied research in today’s interdisciplinary concept of biological sciences.
Learning outcomes
Student will be able to:
-measure absorption, excitation and emission spectra of a fluorophore
-use fluorescence spectroscopy for qualitative and quantitative analysis of biomolecular interactions
- design optimal experimental arrangements using fluorescence spectroscopy and microscopy
Syllabus
  • Spectral characterization of proteins and nucleic acids
  • Accurate determination of concentration of nucleic acids and proteins by fluorescence and absorption spectroscopy (colorimetry)
  • Effects of pH and temperature on spectral properties of fluorophores
  • Measurement of intrinsic fluorescence of BSA, tryptophan, and thyroxin
  • Preparation of fluorescently labeled DNA
  • Spectroscopic characterization of fluorescently labeled DNA
  • Visualization of macromolecules after electrophoretic separation
  • Use of resonance energy transfer (FRET) for detection of DNA hybridization
  • Fluorescence quenching after relaxation of loop structure of fluorescently labeled DNA
  • Real-time PCR – Detection of DNA amplification
  • Fluorescence microscopy (fluorescence in situ hybridization)
  • Fluorescence anisotropy – Study of protein binding to a fluorescently labeled DNA
Literature
  • Lakowicz J.R.: Principles of Fluorescence Spectroscopy. Third Edition, Springer + Business Media, New York, 2006
Teaching methods
laboratory practical sessions
Assessment methods
colloquium
Language of instruction
Czech
Further Comments
Study Materials
The course is taught annually.
The course is taught: in blocks.
Teacher's information
https://is.muni.cz/auth/el/1431/podzim2017/C7230/um/web/index-en.html
The course is also listed under the following terms Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2018, Autumn 2019, Autumn 2020, Autumn 2022, Autumn 2023, Autumn 2024.
  • Enrolment Statistics (autumn 2021, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2021/C7235