Last update: doc. RNDr. Václav Martínek, Ph.D. (30.09.2023)
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.
Literature
Last update: doc. RNDr. Václav Martínek, Ph.D. (16.05.2021)
Manuals for each method and examples of protocols are available in Moodle.
Requirements to the exam -
Last update: RNDr. Jana Rubešová, Ph.D. (05.05.2021)
Credit: successful work in the laboratory on all given laboratory tasks, delivery of the final report in required quality and in time, oral discussion of the report.
Last update: RNDr. Jana Rubešová, Ph.D. (05.05.2021)
Zápočet: úspěšné absolvování zadaných úloh, vypracování přiděleného protokolu v odpovídající kvalitě, jeho včasné odevzdání a diskuse.
Syllabus
Last update: doc. RNDr. Václav Martínek, Ph.D. (30.09.2023)
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
Entry requirements -
Last update: doc. RNDr. Václav Martínek, Ph.D. (30.09.2023)
The laboratory course is suitable for students of the 3rd year of bachelor's studies in biochemistry or molecular biology disciplines. Most tasks are focused on advanced instrumental biochemical methods, so we assume that participants are equipped with knowledge of basic chemical and biochemical techniques in addition to the theoretical understanding of biochemistry. And that they have already successfully completed the introductory "Biochemical Laboratory" courses or equivalent subjects, an entrance test can verify the student's knowledge.
Last update: RNDr. Jana Rubešová, Ph.D. (05.05.2021)
Praktikum je určeno pro studenty 3. ročníku bakalářského studia biochemie. Vzhledem k tomu, že se zaměřuje na pokročilé instrumentální biochemické metody, předpokládáme, že účastníci jsou kromě teoretické znalosti biochemie vybaveni také znalostmi základních chemických a biochemických metod. Tedy že již úspěšně absolvovali Praktikum z laboratorní techniky a také Biochemické praktikum (MC250C31N), popř. ekvivalentní předměty.
