SubjectsSubjects(version: 901)
Course, academic year 2022/2023
  
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 2021
Semester: winter
E-Credits: 5
Examination process: winter s.:
Hours per week, examination: winter s.:0/4 C [hours/week]
Capacity: 48
Min. number of students: unlimited
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): RNDr. Marek Ingr, Ph.D.
Mgr. Petr Jeřábek, Ph.D.
RNDr. Matyáš Krijt, Ph.D.
doc. RNDr. Václav Martínek, Ph.D.
Mgr. Ondřej Skořepa, 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
Opinion survey results   Examination dates   Schedule   
Annotation
Last update: RNDr. Jana Rubešová, Ph.D. (05.05.2021)
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, gel permeation and affinity chromatography, chromatography on reversed phase), electromigration methods (agarose, SDS polyacrylamide and native electrophoresis for separation and visualization of DNA or 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, next-generation DNA sequencing), 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 ("defence").

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.

Syllabus
Last update: doc. RNDr. Václav Martínek, Ph.D. (09.11.2021)

Tasks:

  • IEX - Separation of holo- and apoform of cytochrome b5 by ion exchange chromatography and gel filtration
  • IMAC & WB - Isolation, purification, electrotransfer and immunodetection of TEV protease and 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
  • PYM - Visualization of biomacromolecules in PyMol software
  • 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. (14.02.2022)
The internship is intended for students in the 3rd year of bachelor's studies in biochemistry and international students. Given that it focuses on advanced instrumental biochemical methods, we assume that participants are equipped with knowledge of basic chemical and biochemical techniques in addition to the theoretical understanding of biochemistry. We assume that participants have already completed the Laboratory Technology Laboratory and the Biochemical Laboratory (MC250C31N) or equivalent subjects.
 
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