87818 - Applied Biochemistry

Course Unit Page


This teaching activity contributes to the achievement of the Sustainable Development Goals of the UN 2030 Agenda.

Good health and well-being

Academic Year 2021/2022

Learning outcomes

Attending this course Students will learn the general structure of nucleic acids and proteins, their significant cellular functions and the main protocols and technologies in use for their analysis in a wetlab. Students should finally -understand relevant issues when working in a molecular biowetlab, - proficiently communicate and interact with life scientists.

Course contents

- Setting a research protocol: instruments and reagents in a biomedical laboratory.
- Setting a research protocol: cell cultures.
- Imaging and counting cells: principles of optical microscopy (bright field, phase contrast).
- Molecular components of a cell: aminoacids and proteins in solution.
- UV and visible spectroscopy to rapidly determine nucleic acid or protein concentrations.
- Resolving proteins in a mix: polyacrylamide gel electrophoresis (PAGE) of proteins, two-dimensional electrophoresis of proteins.
- Western blotting.
- Antibodies as laboratory reagents (mono- polyclonal immunoglobulins: sensitivity vs. specificity; species-specificity.
- Antibody functionalization (immunoenzymatic assays, immunofluorescence assays).
- Signal processing in a biomedical laboratory: threshold, dynamic range, resolution, saturation, reference systems.
- Enzyme Linked Immunosorbent Assay (ELISA).
- 2-D Fluorescence Difference Gel Electrophoresis (DIGE).
- Molecular components of a cell: nucleotides and nucleic acids in solution.
- Flow of genetic information.
- DNA replication.
- Oligonucleotide probes in a biomediacl laboratory - how to design them.
- Polymerase Chain Reaction (PCR).
- DNA sequencing.
- Southern blotting - Restriction Fragment Length Polymorphism (RFLP).
- Northern blotting vs. RT-PCR.
- Real-time RT-PCR (qPCR - Livak; primer efficiency; amplicon specificity).
- DNA microarrays to measure gene expression levels.


The slides presented in classes will be available @ IOL.

Wilson and Walker's principles and techniques of biochemistry and molecular biology
by Wilson, Keith | Walker, John M. | Hofmann, Andreas| Clokie, Samuel [2018 edition]. | Cambridge : Cambridge University Press, 2018.
ISBN 978-1-107-16227-3 Hardback
ISBN 978-1-316-61476-1 Paperback

is the textbook for this course.

Teaching methods

Classes. Practical activity for small student's groups in the lab.

Assessment methods

The path of the life sciences has ventured along a cycle of observations, proposals of hypotheses and their experimental verifications, which have become increasingly sophisticated as the instrumental support in the laboratory has allowed it. For the success of this operational strategy it was necessary that the intuition of brilliant scientists was combined with the expertise of brilliant engineers. The terrain of their confrontation must necessarily be a platform of shared communication. And this course aims to offer engineers ideas to open a dialogue with those who work in the life sciences domain.

The verification of learning takes place through a single oral test (typically lasting about 45 minutes) in which a topic drawn by lot [(e.g. liquid chromatography; DNA sequencing; Southern blot (RFLP); northern blot; western blot; polymerase chain reaction; Bradford assay (vis spectroscopy); Enzyme-Linked ImmunoSorbent Assay (ELISA); lateral flow assays; DNA microarrays; protein electrophoresis; nucleic acid quantification (UV spectroscopy)] will be addressed. The examiner will ask questions aimed at ascertaining the knowledge of the specific protocol and inherent technological aspects presented in class. Solutions to practical problems typically emerging during the implementation of the specific protocols, also highlighted during the hours spent in the classroom, may also be required.

Therefore, the exam is an oral test, but it is not a tale where memory prevails over logics. Rather, the examiner will evaluate how much the contents developed in the classroom are effective starting points for operational reasoning. Understanding and applying is the appropriate style for learning verification of this course. In particular, an engineer is required to certify the robustness of the measurement of biomolecular phenomena through the reliability of the signals obtained in the laboratory. The management of issues related to analytical protocols and aspects (e.g. amplification, filtering, sensitivity, specificity, resolution, threshold, dynamic range, saturation...) related to the respective signals that can be acquired must therefore be clear to students. Obviously the frequency of lessons, theoretical and practical, is of great help for this purpose, although it is not a mandatory requirement.

An organic vision of the topics addressed in class, the presence of a critical sense applied to analytical needs, the possession of clarity and expressive precision and specific language will be evaluated with marks of excellence.

Having studied but uncritically expressing the contents, communicating them in a non-specialist form will lead to proportionally less remunerative scores.

You do not pass the exam in case of training gaps.

Teaching tools

Practical activity for small student's groups in the Mol Cell Engn Lab "Silvio Cavalcanti" #3014.

Specific protocols will also be implemented in the classroom.

Office hours

See the website of Emanuele Domenico Giordano