Biomedical Laboratory techniques

Definition and scopes of Clinical Molecular Biology in diagnosis. Small scale mutations, point mutations, repeat expansion mutations and large-scale mutations. Basic molecular methods necessary to diagnostic molecular analyses of nucleic acids. Isolation of nucleic acids. Restriction enzymes and their use in DNA sequence analysis. Southern and Northern blotting. PCR-based techniques (PCR, RT-PCR and quantitative PCR). Molecular Biology in the diagnosis of genetic diseases. Direct and indirect genetic testing. Analysis of DNA polymorphisms. Sequence (SNPs, RFLP) and length (VNTR, STR) DNA polymorphisms. Methods for the detection of known mutations (ASO, reverse dot-blot, ARMS-PCR). Methods for searching for unknown mutations (DGGE, ASO, SSCP, DNA sequencing). Examples of molecular tests used to diagnose and monitor specific genetic diseases (cystic fibrosis, hemoglobinopathies, muscular dystrophies, fragile X syndrome). Forensic applications of molecular biology.

Lecture notes Kaplan JC., Delpech M. Biologia molecolare e medicina. Idelson-Gnocchi. Strachan T., Read A. Genetica molecolare umana. Zanichelli. Molecular Diagnostics: For the Clinical Laboratorian (2nd edition) by William B. Coleman and Gregory J. Tsongalis. Humana Press