The Molecular Biology laboratory operates in the field of translational research dealing with the clinical validation of both protein and molecular markers in neoplastic and non-neoplastic diseases.
The activity involves the extraction of nucleic acids (DNA and RNA) and proteins and subsequent analysis by different molecular approaches (qualitative and quantitative PCR , sequencing, etc.) using conventional methods of analysis (in situ hybridization, direct sequencing, pyrosequencing, real time PCR) and more advanced Next Generation Sequencing techniques as well as fluorescent probe in situ hybridization (FISH) methods.
The Laboratory carries out research and care activities for specialized fields, performing molecular investigations for diagnostic, prognostic and predictive purposes of response, understood as responsiveness/resistance to new oncological drugs (target therapy).
All molecular investigations performed in the laboratory take into account specific protocols defined at the national level by the scientific societies of medical oncologists (AIOM) and anatomic pathologists (SIAPEC-IAP). The laboratory's activities have been validated over time through participation in Italian quality controls organized by scientific societies (AIOM-SIAPEC) and regional (CQ-RER) and in European quality control programs (EMQN).
The Laboratory offers the possibility of traineeships and internships to students, residents of the University and support to doctoral students engaged in research activities in the medical field.
Main lines of research:
The main objective of the research activity is translational and consists mainly in deepening the knowledge of molecular mechanisms involved in oncological pathologies starting from the understanding of the information contained in the genome of cancer cells. In particular, analyses are aimed at observing how changes in the sequence, structure, and functional state of the genome influence the development and progression of tumors. The goal is to identify new molecules associated with neoplasms that can serve as markers or therapeutic targets. The most impactful applications involve making early diagnosis of neoplasia, identifying markers that predict patient response to new targeted drugs. In particular, biological drug resistance studies are performed in lung , breast, brain, gastrointestinal, and lymphoma cancers.
The laboratory constantly collaborates with various study groups from both our University and University Hospital Company and with other regional and national centers and pharmaceutical industries.
Techniques in use
- Nucleic acid extraction
- StandardPCR
- Real timePCR
- Gel electrophoresis
- Fragment analysis by automatic sequencer in capillary
- Pirosequencing
- Sequencing by Next Generation Sequencing (NGS) Platform Ion Torrent™ S5
- Analysis of transcripts by Nanostring technology
- Analysis of fusion transcripts by RT PCR
- Study of epigenetic modifications
- Study of polyclonality
- Analysis of chromosomal alterations with probes by fluorescent in situ hybridization (FISH) In situ hybridization for miRNA
- In situ hybridization by RNAscope technology
Main instrumentation provided
Instrumentation in the molecular Biology laboratory includes machines for PCR and RT-PCR, automatic extractor for nucleic acid extraction and purification, electrophoresis chambers , spectrophotometers, hybridizer for FISH, small laboratory instrumentation. The laboratory is equipped with highly innovative instrumentation including fluorescence microscope with image analysis system for the acquisition and evaluation of chromosomal alterations on interphase nuclei; Advanced Platforms for massively parallel sequencing of DNA/RNA, fundamental in oncology diagnostics, medical genetics and research; Nanostring Platform that studies gene expression and allows the evaluation of its variation in more than 700 genes simultaneously per single sample and the analysis of molecular alterations such as fusions and translocations.
The list of major major instrumentations follows:
- RotorGene Q MDx - Real-Time PCR and accessories
- Thermal Cycler AB 2700 - Thermal Cycler
- Thermal Cycler Applied Biosystems AB 2720 - Thermal Cycler
- EasyPgx pPCR Instrument 96 -Aria Real-Time PCR System
- Ion Torrent™ Next-Generation Sequencing
- Ion One Touch Es
- Ion One Touch 2
- Miseq Illumina System
- Nanostring-nCounter Instrument System (Prep Station, Digital Analyzer)
- Fluorescence microscope with image analysis system (NIS Elements AR)
- Thermobrite
- Qubit4_Fluorometer fluorometer
