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.
Poslední úprava: Martínek Václav, doc. RNDr., Ph.D. (30.09.2023)
Literatura - angličtina
Manuals for each method and examples of protocols are available in Moodle.
Poslední úprava: Martínek Václav, doc. RNDr., Ph.D. (16.05.2021)
Požadavky ke zkoušce -
Zápočet: úspěšné absolvování zadaných úloh, vypracování přiděleného protokolu v odpovídající kvalitě, jeho včasné odevzdání a diskuse.
Poslední úprava: Rubešová Jana, RNDr., Ph.D. (05.05.2021)
To obtain credit: successful work in the laboratory on all given laboratory tasks, submission of the final report in the required quality and time, oral discussion of the report is required.
Note: The grade cannot be entered into the university's student information system, but upon request we can issue an official letter stating the grade.
Poslední úprava: Martínek Václav, doc. RNDr., Ph.D. (29.01.2025)
Sylabus - angličtina
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
Poslední úprava: Martínek Václav, doc. RNDr., Ph.D. (30.09.2023)
Výsledky učení -
Student využívá základní bioinformatické techniky ke studiu biomolekul a provádí vizualizace biomolekul v programu PyMol.
Student provádí separaci vybraných proteinů pomocí iontově výměnné chromatografie na přístroji FPLC.
Student izoluje a purifikuje rekombinantní proteiny, jako jsou TEV proteasy nebo zelený fluorescenční protein.
Student provádí krystalizaci proteinů a má přehled v metodách studia struktury biomakromolekul.
Student provádí polymerasovou řetězovou reakci (PCR).
Student separuje rostlinné pigmenty pomocí chromatografie na reverzní fázi (HPLC).
Student využívá citační manažer a pracuje s odbornou literaturou.
Student zpracuje výsledky úlohy do formy krátké závěrečné odborné práce.
Poslední úprava: Martínek Václav, doc. RNDr., Ph.D. (30.09.2024)
The student uses basic bioinformatics techniques to study biomolecules and performs visualizations of biomolecules in PyMol.
The student performs separation of selected proteins using ion exchange chromatography on an FPLC instrument.
The student isolates and purifies recombinant proteins such as TEV proteases or green fluorescent protein.
The student performs protein crystallization and is familiar with methods for studying the structure of biomacromolecules.
The student performs the polymerase chain reaction (PCR).
The student separates plant pigments using reversed-phase chromatography (HPLC).
The student uses a citation manager and works with the literature.
The student compiles the results of the assignment into a short final research paper.
Poslední úprava: Martínek Václav, doc. RNDr., Ph.D. (30.09.2024)
Vstupní požadavky -
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.
Poslední úprava: Rubešová Jana, RNDr., Ph.D. (05.05.2021)
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.
Poslední úprava: Martínek Václav, doc. RNDr., Ph.D. (30.09.2023)