66435 - Applied Molecular Biology

Learning outcomes

The students will be able to operate by ourself in a clinical-pathology laboratory. Furthermore, they will be able to effectively join a lab team, to organize the diagnostic procedures and to perform clinical and molecular assays.

Course contents

-- The course of APPLIED MOLECULAR BIOLOGY is part of the integrated course ATTIVITA' SPERIMENTALE NEL LABORATORIO BIOTECNOLOGICO (C.I.). At the end of the integrated course the student is able to plan and undertake research in animal biotechnology. The student knows the molecular basis of biological processes applied to cell cultures and animal models, as well as the rules of experimental design in compliance with legal regulations. The student is able to perform cell cultures and analyze nucleic acids and proteins. The verbalizing teacher for the integrated course is Prof. Monica Forni.

The course of APPLIED MOLECULAR BIOLOGY, incuding three modules, aims to teach students the basics of the main techniques of molecular biology. The contents of module 1 (2 CFU) are related to acquire the theoretical basis of the main proteomics techniques (extraction, protein quantification, sub-protein separation, single and two-dimensional electrophoresis) and the theoretical basis of genomics applied to forensic and conservation genetics.

Module 2 is related to: basic techniques of molecular biology (DNA isolation, Polymerase Chain Reaction, Sanger sequencing, NGS sequencing, analysis of fragments, single nucleotide polymorphisms (SNPs), basic techniques of Molecular diagnostics, authomated and/or manual clinical techniques, in particular urinalysis, serum protein electrophoresis, serum chemistry analysis. In particular, to learn methods for performing molecular assay aimed at diagnosing neoplastic and genetic diseases as well as genotyping microsatellites for parentage verification in domestic animals. To learn skills for performing automated and manual clinico-pathological analysis and in particular clinical-chemistry, hematology, electrophoresis, urinalysis, fecal examination, hematological and cytological staining, coagulation assay.  For Molecular diagnosis: 1.Genetics of immunoglobulins; definition of lymphoid marker and Ig and TCR gene rearrangements. Heteroduplex analysis and fragment analysis as diagnostic method for detecting Ig and TCR clonal rearrangements. Methods for purifying DNA for the molecular diagnosis. 2.Role of somatic mutations in cancerogenesis. “Drivers” and “passengers” mutations. Techniques used for the highly sensitive detections of somatic mutations in neoplastic tissues. 3.Repeated DNA sequences. Microsatellites as polymorphic markers for parentage testing and pedigree verification. 4. Techniques for genotyping single nucleotide variations: restriction enzymes, real-time PCR, High resolution melting curve analysis, allele-specific primer extension methods.

Module 3 has a strictly practical approach and provides training based mainly on laboratory activities. Each exercise will be introduced by a short theoretical lesson to contextualize the purpose of the analysis and the student will carry out the practical activities individually, applying a protocol that will be provided and explained until the results are obtained through the applied analytical technique also through the processing of instrumental data. The student will learn in particular techniques of: extraction, purification, separation, quantification and qualitative analysis of proteins by means of gelfiltration chromatography, quantitative colorimetric determinations and 1D electrophoresis, which will be applied to different biological matrices. Students who are not present will be able to learn remotely the methods carried out with the help of specially prepared videos or available on the web.

 

 

Readings/Bibliography

Powerpoint lectures will introduce the practical sessions. For some cases, brief videos will be used to illustrate the techniques.

Teaching material can be found at: Virtuale (unibo.it) [https://virtuale.unibo.it/]

Proteomica T. Alberio, M. Fasano, P. Roncada (EdiSES, 2021)

Teaching methods

For practical work, groups composed by 2 or 3 students will perform the molecular assays. Innovative teaching techniques will be applied, aimed at obtaining an active participation of the students.

It is mandatory to wear a gown for access to the teaching laboratory. When necessary, PPEs will also be provided for carrying out specific activities, such as single-use gloves and safety goggles.

Students connected remotely will be guaranteed the opportunity to watch live coverage of all the practical activities that are carried out in order to understand how the individual steps of the different methods are carried out.

Considering the type of activities and teaching methods adopted, the attendance of the training activities of modules 2 and 3 requires the performance of modules 1 and 2 in e-learning mode of specific training on safety and health in the workplace [https://www.unibo.it/it/servizi-e-opportunita/salute-e-assistenza/salute-e-sicurezza/sicurezza-e-salute-nei-luoghi-di-studio-e-tirocinio] and the participation in module 3. Information on dates and modalities of attendance of module 3 are available in the appropriate section of the course website.

As far as module 3 is concerned, in order to facilitate the learning process, each student will be given a handout with the protocols of the practical activities explained; the handout will also contain worksheets that each student will have to complete by collecting the data obtained from his/her experimental test; at the end of the practical activity students will have to be able to process the data collected with appropriate spreadsheets and discuss the final results in group work.

Please note that in case of emergency conditions for Covid-19, teaching activities may undergone variations. In particular, the clinical-pathology lab cannot host the practical activities. The clinical-pathology procedures will be illustrated using videos and other multimedia tools and the Teams platform remotely available.

Students with special needs are invited to contact the teacher by e-mail to illustrate them and to organize the teaching and examinations in the most suitable way.

Assessment methods

The evaluation is expressed by a grade in thirtieths. The test is passed with a grade of at least 18/30. In case of the highest grade (30/30), honors may be awarded. The student has the right to refuse the verbalization of the proposed positive grade at least once. Student who fails the exam may take it again three weeks after the previous attempt.

The final grade refers to the achievement of educational objectives:

- preparation concerning a limited number of fundamental topics and ability to analyze that emerges only with the help of the teacher, overall correct language: 18-20;

- preparation on a limited number of fundamental topics covered in the course and ability to analyze the fundamental issues of metabolism and transmission of information, correct language: 21-24;

- preparation on a large number of topics, ability to make autonomous choices of critical analysis, mastery of specific terminology: 25-29;

- comprehensive preparation on all topics covered in the course, ability to make independent choices of critical analysis, deep knowledge of specific terminology: 30-30L.

The exam consists of a written test containing open and multiple choice questions, which can be followed by an oral test. The exam is intended to evaluate the acquisition of knowledge and skills as reported in the course program.

This class is part of an Integrated Course, the final grade will be proportional to the CFU of each teaching

35106 Policy and Legislation in Animal Biotechnology 3CFU 20%

66437 Applied Experimental Biology 6CFU 40%

66435 Applied Molecular Biology 6CFU 40%

It should be noted that if the emergency conditions for Covid-19 continue, the modalities of the final test may change

Exam reservation and dates of exam sessions - Students can make a reservation of the exam at: (https://almaesami.unibo.it/almaesami/welcome.htm ). Studends that have regularly attended the course can take the exam at fixed dates within specific periods of the academic year.

For special educational needs, students with disabilities or DSAs can contact the faculty member by email by entering the appropriate University service reference personnel in CC.

Teaching tools

Power point presentations, virtual labs, databases, practical laboratory work.

During the lessons digital tools will be used to support and integrate lectures and specific apps (Kahoot!) for conducting surveys and questioning sessions.

Students attending in presence will have single workstations equipped with laboratory equipment suitable for carrying out the practical activities presented. For students attending remotely will be made video recordings during the execution of the methods. For all students power point presentations will be used for common explanations of the different protocols and excel spreadsheets for demonstrations of data processing methods.

Students with special needs are invited to contact the teacher by e-mail to illustrate them and to organize the teaching and examinations in the most suitable way.

Office hours

See the website of Gloria Isani

See the website of Fabio Gentilini

See the website of Giulia Andreani