Molecular analytical methods - TRASCRITTOMICA (2013/2014)

Course code
4S02774
Credits
4
Academic sector
CHIM/01 - ANALYTICAL CHEMISTRY
Language of instruction
Italian
Teaching is organised as follows:
Activity Credits Period Academic staff Timetable
teoria 1 II semestre Diana Bellin
laboratorio 3 II semestre Diana Bellin

Lesson timetable

II semestre
Activity Day Time Type Place Note
teoria Monday 2:30 PM - 4:30 PM lesson Lecture Hall L from Apr 7, 2014  to Jun 13, 2014
laboratorio Monday 2:30 PM - 7:30 PM lesson Laboratory Laboratorio di Biotecnologie genetiche  

Learning outcomes

This course introduces to some new analytical technologies widely used for genome analysis, functional genomics and trascriptomics. The purpose is to provide both knowledge of theory for these technologies as well as practical experience.

Syllabus

THEORY:
GENOMICS:
-GATEWAY TECHNOLOGY: Lamba phage recombination and introduction to gateway technology, BP and LR reactions, ccdB based selection, ENTRY clone production, TOPO cloning, gateway compatible destination vectors.
-MUTAGENESIS: DNA mutations, mutagenesis in forward and inverse genetics, random mutagenesis (chemical or PCR based), site directed mutagenesis (with or without PCR), Quickchange, DNA shuffling
TRANSCRIPTOMICS:
-RT-PCR AND REAL TIME RT-PCR: single gene expression analysis and genome-wide expression analysis, RT-PCR, Real Time RT-PCR, absolute and relative quantification (delta delta Ct method for data analysis), SYBR green analysis, TaqMan probes, LUX, Molecular Beacons and Scorpions
-MICROARRAY: cDNA microarray and their production, oligoarray and oligoarray production, direct and indirect probe labeling, experimental design and controls for microarray experiment setup, hybridization, image acquisition and data analysis

PRACTICAL PART:

Gene Cloning using the gateway technology and transfer of a gene in a destination vector
Expression analysis using real time RT-PCR
Microarray expression analysis

Assessment methods and criteria

See main page of the integrated course