Biochemical Techniques for The Analysis Laboratory

A.Y. 2023/2024
3
Max ECTS
32
Overall hours
SSD
BIO/10
Language
Italian
Learning objectives
Provide the theoretical-practical knowledge of basic laboratory techniques for biochemical analyzes aimed at the qualitative and quantitative analysis of biological samples / foods / materials by spectrofluorimetry / colorimetry; provide knowledge of the foundations and main applications of analytical and preparatory biochemical techniques based on electrophoresis and chromatography
Expected learning outcomes
Knowledge and understanding: theoretical/practical principles of basic biochemical techniques aimed at the qualitative and quantitative analysis of biological samples/food/materials by spectrofluorimetry/colorimetry and analytical and preparative techniques based on electrophoresis and chromatography
Applicative skills acquired: preparation and execution of simple assays based on colorimetry/fluorescence
Communication skills and independent judgement: preparation of simple reports for the communication of the results obtained in the tests performed in the laboratory / evaluation of their reliability
Single course

This course can be attended as a single course.

Course syllabus and organization

Single session

Responsible
Lesson period
First semester
Course syllabus
Frontal teaching
Safety procedures in a biochemical laboratory (0.5 h)
Description of the main solvents used in biochemical laboratories (0.5 h)
Review of general chemistry topics related to units of measurement of the quantity and concentration of species in solution (1 h)
Buffer solutions used in the laboratory; buffer systems in living organisms (2 h)
Principles of spectrophotometry; use of UV-visible spectrophotometry for the dosage of molecules from biological matrices and foods, including enzymes (5 h)
Principles of spectrfluorometry (1 h)
Principles of electrophoresis applied to protein analysis (3 h)
Chromatographic techniques: principles and applications for the purification and analysis of compounds (3h)

Exercise teaching
Acid/base titration and preparation of buffer solutions (4 h)
UV-visible spectrophotometry and colorimetry: determination of the UV-vis absorption spectrum of a compound, assay of molecules by colorimetry, setting up and carrying out an experiment for the determination of the protein content of a sample using a calibration curve, enzyme assays (5 hours)
Electrophoresis of proteins on polyacrylamide gels in the presence of sodium dodecyl sulfate (4 h)
Preparation of a chromatographic column and its use for the separation of compounds by gel filtration (3 h)
Prerequisites for admission
Topics of general chemistry provided by previously attended Chemistry courses relating to definitions of pH, buffer systems, concentration, nomenclature of the main inorganic and organic compounds useful for the preparation of aqueous buffers; concepts acquired in previously attended Biochemistry courses related to enzymes, substrates, inhibitors and their biological relevance
Teaching methods
Lectures on theoretical aspects of the methodologies object of the subsequent practical exercises
Experimental procedures carried out partly in the teaching laboratories and partly in the research laboratories of the biochemical group
Teaching Resources
The reference material will be provided by the teacher through the new MyARIEL platform soon to be introduced. The relevant link will therefore be communicated directly to the students
Assessment methods and Criteria
Written exam lasting a total of 1.5 hours consisting of 1 open question on the topics covered by the course and a graph and/or numerical exercise for the calculation of experimentally measurable parameters starting from data provided by the teacher. The final overall vote will be calculated as the average of the votes obtained in the two parts of the exam. In particular, it will take into account the completeness, clarity and correctness of the contents for the answer to the open question and the correctness of the numerical result obtained and the clarity and completeness of the elements of the graph that may be required to complete the answer to the second question.
BIO/10 - BIOCHEMISTRY - University credits: 3
Practicals: 16 hours
Lessons: 16 hours
Professor: Negri Armando