Teacher
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PROIETTI Silvia
(syllabus)
Theoretical part (32 hours) Introduction to nucleic acids and proteins handling: Methods for DNA, RNA and proteins isolation from biological samples and related analysis. Comparison between protein samples: differential-in-gel electrophoresis (DIGE). Methods for gene expression analysis: RT-PCR. DNA microarray technology. RNA-Seq. Methods for protein-protein interaction analysis: Far-Western, Pull-down, yeast-two/three hybrid assay, Co-Immunoprecipitation, Tandem Affinity Purification (TAP) system, Phage Display, Bimolecular Fluorescent Complementation (BiFC), FRET. Methods for DNA/RNA-protein interaction analysis: Chromatin Immunoprecipitation assay (ChIP and ChIP-on-chip). Electrophoretic Mobility Shift Assay (EMSA). DNA pull-down. Southwestern. Yeast three-hybrid system. Methods for epigenetic modifications analysis : DNA Methylation analysis: Methylation-Sensitive Amplification Polymorphism (MSAP). Bisulfite (non methylation)-specific PCR and Methylation-specific PCR (MSP). Methods for histone modifications analysis by ChiP. Mutagenesis techniques and genome editing: Site-directed mutagenesis and CRISPR-Cas9 System. Diagnostic application of PCR. Next Generation Sequencing: second and third generation sequencing platforms.
Practical part (16 hours) Total RNA extraction, RT-qPCR. Genomic DNA extraction. DNA methylation analysis. Total protein isolation and protein-protein interaction assays.
(reference books)
Brown T.A. Gene cloning and DNA analysis: an introduction. 7th ed., 2016, Wiley-Blackwell. Watson J.D., Caudy A.A., Myers R.M., Witkowski J.A. Recombinant DNA, genes and genomes – a short course. 3rd ed., 2007, W.H. Freeman & Co. Lesk A.M. Introduction to Genomics. 3rd ed., 2017, Oxford University press.
Slides are available in the teaching platform. Handouts are provided by the teacher for practical activities. Non-attending students are encouraged to contact the teacher for information about the program, teaching materials, and the examination mode.
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