The course aims to provide students with the theoretical and practical basic notions of some of the most important techniques commonly used in the molecular / biotechnological field. For this purpose, both cell biology techniques, with particular reference to the management of cell cultures (human and mouse), and fluorescence analysis techniques, such as fluorescence microscopy and flow cytometry, applied to the study of cell populations will be addressed. Finally, space will also be given to sequencing techniques with particular attention to the most innovative new generation techniques (next-generation sequencing - NGS).

1 - Cell biology techniques

- Safety and sterility in cell culture laboratories

- Primary cells, cell lines, immortalized cell lines

- Practices in the management of cell cultures

- Instrumentation in cell cultures (cell culture hood and incubator)

- Cell culture media and consumables

- Cell counting strategies

- Exercise: thawing, counting, maintenance and freezing of immortalized human cell lines

2 - Fluorescence microscopy techniques

- Fluorescence analysis

- Quantitative and qualitative determination

- The fluorescence microscope

- Exercise: cellular localization of a protein by immunofluorescence

3 - Flow cytometry techniques

- The cytofluorimeter

- Flow cytometry methods

- Exercise: cytofluorimetric analysis of the cycle of tumor cells subjected to treatment with potential anticancer molecules

4 - DNA sequencing techniques

- Sanger technique

- The Sanger sequencer

- Next generation sequencing (NGS)

- NGS sequencers

- Exercise: Sanger sequencing and presentation of the NGS Ion S5 sequencing system

5 - Epigenetic techniques

- DNA methylation analysis

- Chromatin analysis: chromatin immunoprecipitation (ChIP)

None.

Knowledge and understanding - The student must demonstrate to have acquired the theoretical / practical knowledge of the experimental techniques addressed during the course. The student must also demonstrate to be familiar with the safety standards relating to the techniques taken into consideration, as well as those essential in order to avoid possible contamination from external agents or the introduction of variables that could compromise the interpretation of the essay itself. Finally, the student will have to demonstrate to have understood the importance of using experimental controls suitable for the assay and the experimental context.

Ability to apply knowledge and understanding - The student will have to demonstrate to have mastered the techniques covered during the course and to be able to independently evaluate their use according to a specific experimental context, as well as the development of a correct experimental design and the use of appropriate controls. The student must also demonstrate to have acquired specific language properties related to the techniques that will be addressed.

Independent judgment - The student must be able to critically elaborate the notions provided during the course and to independently evaluate the results of the techniques addressed during the course itself.

Communication skills - The student must demonstrate to have command of the language of the subject of the course and to be able to correctly expose the technical and scientific concepts that will be treated.

Learning skills - The student must demonstrate to have acquired the ability to independently find information about the subject of the course by consulting the relevant scientific publications. This ability will be encouraged, as far as possible, by the teacher through specific in-depth studies that the students can carry out independently and which will be discussed in the classroom.

The teaching material prepared by the lecturer in addition to recommended textbooks (such as for instance slides, lecture notes, exercises, bibliography) and communications from the lecturer specific to the course can be found inside the Moodle platform › blended.uniurb.it

Teaching

The course will take place through lectures in which the teacher exposes, through visual aids (slides), the fundamental notions and provides practical examples to facilitate understanding, as well as through laboratory exercises relating to the techniques addressed. Students will be involved during the teaching activity also for the purpose of verifying the effectiveness of the tools used and the understanding of the topics covered, as well as in carrying out the laboratory exercises in which they will independently carry out the proposed activities.

Attendance

Obligation to attend 2/3 of laboratory activities.

Course books

Didactic study material will be provided by the teacher during the course.

Assessment

In order to ascertain the preparation at the end of the course, the student will have to take an oral exam aimed at verifying the understanding of the main aspects concerning the molecular biology / biotechnological techniques addressed during the course. The ability to autonomously develop a specific experimental design and to critically evaluate the results obtained by applying the same techniques will also be evaluated. Particular attention will also be paid to the quality of the student's oral presentation. The teacher will also verify the student's ability to adequately manage his own personal laboratory notebook.

Disabilità e DSA

Le studentesse e gli studenti che hanno registrato la certificazione di disabilità o la certificazione di DSA presso l'Ufficio Inclusione e diritto allo studio, possono chiedere di utilizzare le mappe concettuali (per parole chiave) durante la prova di esame.

A tal fine, è necessario inviare le mappe, due settimane prima dell’appello di esame, alla o al docente del corso, che ne verificherà la coerenza con le indicazioni delle linee guida di ateneo e potrà chiederne la modifica.