87818 - Applied Biochemistry

Academic Year 2023/2024

  • Teaching Mode: Traditional lectures
  • Campus: Cesena
  • Corso: Second cycle degree programme (LM) in Biomedical Engineering (cod. 9266)

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.

Readings/Bibliography

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.

Assessment methods

The exam will be a written test (multiple choice questions, open questions and calculations), intended to restitute contents presented in class. The task must be completed in a maximum of 60'.

An organic vision of the topics addressed in class, the presence of a critical sense, 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 learning gaps.

Teaching tools

Specific protocols will be implemented in the classroom.

Office hours

See the website of Emanuele Domenico Giordano

SDGs

Good health and well-being

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