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

This course is part of the integrated cours ATTIVITA' SPERIMENTALE NEL LABORATORIO BIOTECNOLOGICO (C.I.). This course, incuding tthree modules, is aimed to approach 1) basic protocols of proteomics (extraction, quantification separation of sub-proteomes, 1D and 2Delectrophoresis; 2) basic techniques of molecular biology (DNA isolation, Polymerase Chain Reaction, Sanger sequencing, analysis of fragments, single nucleotide polymorphisms (SNPs); 3) basic techniques of Molecular diagnostics; 4) authomated and/or manual clinical techniques, in particular urinalysis, serum protein electrophoresis, serum chemistry analysis.

In particular, to learn methods for performing molecolar 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.

The course is strictly practical. Practical sessions will be introduced by a short lecture to fournish the background of the aims of the assay. Students will learn 1) the extraction and separation methods for proteins, 2) 1D electrophoresis 3) purification nucleic acids from clinical samples and 4) the methods to amplify the target of neoplastic diseases and to perform allelic discrimination assays. Also the amplification of polymorphic traits as microsatellites will be afforded during the course.

The organization and managment of a clinical-pathology Laboratory as well as le operative instructions on how to perform clinico-pathological assays will be presented at the Veterinary Clinical-pathology laboratory. The students will operate ourselves.

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: https://iol.unibo.it [https://iol.unibo.it/]

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 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

Individual skills will be monitored during practical activity.

The final grade will be composed of the active participation during lecture and laboratory activities and a written exam about lecture topics.

This teaching 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%

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.

Teaching tools

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

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

Office hours

See the website of Gloria Isani

See the website of Fabio Gentilini

See the website of Giulia Andreani