Molecular Biology Laboratory

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Specifications: Lab. specialized in molecular virology and viral enzymology.

Equipment available: 2 Laminar Flow biohazard Hoods, 2 Incubators, 1 Refrigerator (+4/-20 °C) and 1 Freezer (-20 °C), 1 Real-Time PCR instrument, 2 PCR thermocyclers, 1 FPLC, 1 Centrifuge, 2 Refrigerated Centrifuges, 2 Microcentrifuges, 2 Spectrophotometers, 1 Orbital shaker incubator, 1 Hybridiser, 1 Transilluminator,  Gel electrophoresis Apparatus for nucleic acids and proteins, 1 Thermostatic Bath, 1 Termoblock.

Year of acquisition: 2004

Responsibles: Dr. Gabriele Giliberti

Ownership: Department of  Biomedical Sciences and Technologies Section of  General Microbiology and Virology & Microbial Biotechnologies, University of Cagliari

Main applications:

♦ Real-time PCR analysis of HIV-1 replication in ex vivo target cells.

♦ Nucleic acid extraction, RT-PCR and PCR for amplification and sequencing of wild type and drug-resistant viruses for identification of mutations involved in the acquisition of drug-resistance to nucleoside and non-nucleoside inhibitors of HIV-1 and other RNA viruses.

♦ Cloning and expression in Escherichia coli of viral genes coding for enzymes target of selective antiviral chemotherapy (i.e. RNA polymerases).

♦ Cloning of site-directed mutated genes coding for target enzymes in order to define the role of sites potentially involved in drug-resistance.

♦ Expression and purification of viral wild type enzymes and mutants to be used in in vitro enzymatic assays to evaluate antiviral inhibitors activity.