SubjectsSubjects(version: 964)
Course, academic year 2024/2025
   Login via CAS
Advanced Practical Biochemistry - MC250C17E
Title: Advanced Practical Biochemistry
Czech title: Pokročilé praktikum z biochemie
Guaranteed by: Department of Biochemistry (31-250)
Faculty: Faculty of Science
Actual: from 2024
Semester: winter
E-Credits: 5
Examination process: winter s.:
Hours per week, examination: winter s.:0/4, C [HT]
Capacity: 30
Min. number of students: unlimited
4EU+: no
Virtual mobility / capacity: no
State of the course: taught
Language: English
Is provided by: MC250C17A
Explanation: in English
Additional information: https://dl2.cuni.cz/course/search.php?search=MC250C17N
Note: enabled for web enrollment
Guarantor: RNDr. Ondřej Vaněk, Ph.D.
doc. RNDr. Václav Martínek, Ph.D.
Teacher(s): Mgr. Maxmilián Hejl
Mgr. Petr Jeřábek, Ph.D.
doc. RNDr. Václav Martínek, Ph.D.
RNDr. Adam Obr, Ph.D.
RNDr. Ondřej Vaněk, Ph.D.
Class: Vysokokapacitní centrifuga s příslušenstvím
Stolní centrifuga s příslušenstvím
Původní předmět
Co-requisite : {At least one of the following subjects: MC250C42N, MC250C31N, MC250C32, MB150C04, MC250C35 MC250C35N, MC250C32N}
Incompatibility : MC250C03, MC250C03E, MC250C17A, MC250C17N
Is incompatible with: MC250C03E, MC250C17A, MC250C03
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.
Last update: Martínek Václav, doc. RNDr., Ph.D. (30.09.2023)
Literature
Manuals for each method and examples of protocols are available in Moodle.
Last update: Martínek Václav, doc. RNDr., Ph.D. (16.05.2021)
Requirements to the exam -

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: Rubešová Jana, RNDr., Ph.D. (05.05.2021)
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.

-----

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

Last update: Martínek Václav, doc. RNDr., Ph.D. (30.09.2023)
Learning outcomes -
  •   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.
Last update: Martínek Václav, doc. RNDr., Ph.D. (30.09.2024)
Entry requirements -
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: Martínek Václav, doc. RNDr., Ph.D. (30.09.2023)
 
Charles University | Information system of Charles University | http://www.cuni.cz/UKEN-329.html