Practical Biochemistry - MC250C35E

• Anglický název: Practical Biochemistry
• Český název: Biochemické praktikum
• Zajišťuje: Katedra biochemie (31-250)
• Fakulta: Přírodovědecká fakulta
• Platnost: od 2022
• Semestr: oba
• E-Kredity: 4
• Rozsah, examinace: 0/4, Z [HT]
• Počet míst: 72
• Minimální obsazenost: neomezen
• 4EU+: ne
• Virtuální mobilita / počet míst pro virtuální mobilitu: ne
• Stav předmětu: vyučován
• Jazyk výuky: angličtina
• Poznámka: povolen pro zápis po webu; předmět lze zapsat v ZS i LS
• Garant: RNDr. Daniel Kavan, Ph.D.
• Vyučující: RNDr. Kateřina Bělonožníková, Ph.D.; RNDr. Daniel Kavan, Ph.D.
• Třída: Vysokokapacitní centrifuga s příslušenstvím; Stolní centrifuga s příslušenstvím; Původní předmět
• Neslučitelnost : MC250C31N, MC250C35
• Je neslučitelnost pro: MC250C35
• Ve slož. korekvizitě pro: MC250C03, MC250C03E, MC250C17A, MC250C17E

Anotace

Practical course in biochemistry is provided by Daniel Kavan, Veronika Hýsková, Josef Chmelík and Kateřina
Bělonožníková
.
The students are dealing with basic biochemical methods used for protein and nucleic acid isolation and their
characterization (extraction, centrifugation, precipitation, dialysis, protein content determination).
Gel filtration is used for protein purification from a mixture or for relative molecular mass determination. The
students determine the relative molecular mass of proteins by SDS polyacrylamide gel electrophoresis. An
important part of the practical course is dealing with enzymes, determination of Michaelis constant, maximal
reaction rate, pH optimum, activation and inhibition, electrophoretic separation of isoenzymes and their detection
in the gel.

Poslední úprava: Kavan Daniel, RNDr., Ph.D. (31.01.2022)

Literatura

Laboratory tasks manual provided as pdf file.

Poslední úprava: Čermáková Michaela, RNDr., Ph.D. (19.04.2021)

Požadavky ke zkoušce

Work on the laboratory task will be allowed only to students who demonstrate a sufficient understanding of the principles of tasks.

It is necessary to consult obtained results with pedagogical staff before leaving the laboratory.

The protocol must be submitted no later than the beginning of the next practice, otherwise the task will be forfeited.

It is advisable to send the electronic version of the protocol by email at least 24 hours before the next laboratory practice (pdf format).

In case of corrections, the protocol must be approved without mistakes within three weeks of the task being elaborated.

At most one approved protocol may be missing at the time of the credit test.

To obtain the credit it is necessary to:

• Experimentally perform specified tasks and report the results

• To present the selected task (in groups)

• To get enough points (2/3) from protocols and final test

• Return the key from the locker in the hallway

Poslední úprava: Čermáková Michaela, RNDr., Ph.D. (19.04.2021)

Sylabus

1. Isolation of a protein - enzyme, determination of specific activity

2. Gel chromatography of proteins (separation, relative molecular mass

determination)

3. SDS - polyacrylamide electrophoresis (determination of relative

molecular mass of proteins)

4. Practical enzymology - kinetic characteristics, Michaelis constants,

maximal reaction rate, pH optimum, activation and inhibition of enzymes,

electrophoretic separation of isoenzymes and detection of the enzyme

activity in gel

5. DNA manipulation (restriction analysis)

6. Oral presentation of one experiment

Poslední úprava: Čermáková Michaela, RNDr., Ph.D. (19.04.2021)

Výsledky učení

The student will apply one of the protein isolation techniques, determine enzyme activity, measure protein concentration, and calculate specific activity.
The student observes the affinity of the enzyme for the substrate and determines Michaelis constant and maximum reaction rate.
The student determines the temperature and pH optimum of a selected enzyme, observes the effect of an inhibitor on enzyme activity, and observes the activity and individual isoforms of a selected enzyme after electrophoretic separation under native conditions.
The student uses gel chromatography to separate proteins, develops a calibration dependence of molecular weight on elution volume, determines the molecular weight of an unknown protein, and determines the distribution coefficient of a selected protein.
The student will perform electrophoretic separation of proteins in a polyacrylamide gel under SDS conditions and determine their relative molecular weight.
The student will measure DNA concentration, perform electrophoretic separation of DNA after restriction endonuclease cleavage, and compare suspect DNA samples to the crime scene.

Poslední úprava: Kavan Daniel, RNDr., Ph.D. (17.12.2024)