|
Lecturer(s)
|
-
Špryncová Martina, Ing. Bc. Ph.D.
-
Štěpánková Šárka, Mgr. Ph.D.
|
|
Course content
|
Safety of work in biochemical laboratory, theoretical introduction - spectrophotometry (absorbance, transmittance, absorption maximum). Techniques of pipetting with piston micropipettes, verification of work accuracy, determination of absorption maximum, calibration, single standard method. Buffer preparation, pH calculation, pH measurement, buffer capacity dependence on buffer concentration, influence of ionic strength on pH buffer. Aminoacids and proteins - biuret reaction, determination of albumin concentration using biuret reaction. Determination of protein concentration by BCA or Bradford method, determination of protein concentration and purity by UV absorbance measurement. Protein precipitation - reversible protein precipitation, protein precipitation associated with denaturation, determination of efficacy of selected deproteinants. Determination of isoelectric point of selected proteins. Qualitative determination of carbohydrates (evidence reactions). Quantitative determination of glucose (enzymatic reaction - glucose oxidase). Vitamins - qualitative determination of vitamin A, D, semiquantitative determination of vitamin C. Nucleic acids - DNA and RNA resolution, DNA and RNA determination, UV absorbance measurement. Credit test.
|
|
Learning activities and teaching methods
|
|
Laboratory work
|
|
Learning outcomes
|
The aim is to improve practical competency of student for application measuring physical and chemical properties of substrates, enzymes, vitamins, hormones and nucleic acids.
Student will be able to work in biochemical laboratories and work with common analytical equipment (centrifugation, extraction, protein precipitation, spectrophotometric measurements). Student can statistically interpret obtained results.
|
|
Prerequisites
|
unspecified
|
|
Assessment methods and criteria
|
Written examination
Compulsory attendance at laboratory exercises. Credit test.
|
|
Recommended literature
|
-
studijní materiály na IS STAG.
-
Štěpánková Š., Královcová P., Kanďár R. Laboratorní cvičení z obecné a klinické biochemie. ISBN 978-80-7395-958-6.
-
Štěpánková Š., Žáková P., Kanďár R. Laboratorní cvičení z obecné a klinické biochemie (třetí doplněné vydání). Pardubice. 2022.
|