Syllabus
Tasks:
INF/PYM - Basics of bioinformatics and Visualization of biomacromolecules in PyMol software
IEX - Separation of selected proteins by ion exchange chromatography on FPLC
IMAC & WB - Isolation, purification, electrotransfer and immunodetection of recombinant protein (e.g. TEV protease or green fluorescent protein)
X - protein crystallization and methods of biomacromolecules' structure determination
PCR - Polymerase chain reaction of DNA
HPLC - Separation of plant pigments by reversed phase chromatography
LIT - Training in work with scientific literature
The course is finished by delivering a short report from one given lab task in a form of short scientific writing.
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The course was innovated within the project ESF for University II at Charles University, reg. No .: CZ.02.2.69/0.0/0.0/18_056/0013322
Annotation
The practical course runs on both days in one week, i.e. both on Mondays and Tuesdays. The task schedule is available upon registration in Moodle and will be created after the first introductory meeting at the beginning of the course where students pass and sign the safety training and also divide into working groups (two people) - the personal attendance at this meeting is mandatory!
The aim of this practical course is to introduce students to both common and modern instrumental biochemical methods, such as extraction of compounds from natural material, UV/VIS absorption spectrophotometry, chromatographic techniques (HPLC, FPLC, ion exchange, hydrophobic and affinity chromatography, chromatography on reversed phase), electromigration methods (agarose and SDS polyacrylamide electrophoresis for separation and visualization of DNA and proteins, Western blot - electroblotting of proteins with their immunodetection), crystallization of proteins and methods of their structural determination, PCR amplification of DNA and related techniques of work with DNA (cloning, real-time PCR resp. quantitative Q-PCR), as well as practical session in computer room devoted to basics of bioinformatics and work with PyMol software for visualization of the structure of biomacromolecules.
The course is finished by another session on searching in biochemical information sources, work with scientific literature and evaluation of experimental results, introduction to principles of text structuring and formal requirements relevant to scientific literature (i.e. bachelor thesis or scientific publications) and subsequent writing of a final report for one given practical task in a form of small scientific work, its timely submission and oral discussion.
Theoretical introduction to all used techniques, supervision of the exercises and discussion of the obtained results is provided in English to all Erasmus students. The written final report is accepted in English.