84222 - Applied biochemistry (2nd cycle)

Academic Year 2018/2019

  • Teaching Mode: Traditional lectures
  • Campus: Cesena
  • Corso: Second cycle degree programme (LM) in Biomedical Engineering (cod. 9243)

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 1) understand relevant issues when working in a molecular biowetlab, 2) have built up a communication platfom to interact with life molecular 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.

Principles and techniques of biochemistry and molecular biology (7th edition, 2010) K. Wilson & J. Walker. Published by Cambridge University Press.

Teaching methods

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

Assessment methods

Oral exam.

Teaching tools

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

Specific protocols will also be implemented in the classroom.

Office hours

See the website of Emanuele Domenico Giordano