Activity | Credits | Period | Academic staff | Timetable |
---|---|---|---|---|
Teoria | 2 | 2nd Semester | Massimo Delledonne | |
Laboratorio | 6 | 2nd Semester | Diana Bellin, Sara Zenoni |
This course aimed to study in detail the manipulation of nucleic acids and the PCR analysis that are at the basis of many methods for molecular analysis.
Theory
Vectors, Cosmids, Fage vectors, Fosmids, YAC e BAC
Vector selection, selectable markers, cloning site
Miniprep
Purification and separation of nucleic acids
Purification of genomic DNA
Purification of RNA
Electrophoresis separation
Manipulation of DNA and cloning
Cloning vectors
E.coli system
Restriction by enzymes
Purification of DNA fragments from gel of agarose
Defosphorilation
Ligation
Competent cells preparation
Transformation
Screening of recombinant clones
PCR
Description of PCR cycle, PCR on genomic DNA, RT-PCR, retrotranscription one-step and two-steps
Molecular Hybridization
Southern Blot
Northern Blot
Laboratory
Isolation of plasmid DNA, genomic DNA and RNA
Separation of nucleic acids by Electrophoresis
Cloning:
Restriction with enzymes
Purification of DNA fragments from agarose gel and from solution
Vector dephosphorilation
Ligation
Competent cells preparation
Transformation and screening of recombinants by colony PCR
PCR on genomic DNA
RT-PCR: quantification, DNase treatment, cDNA synthesis and PCR on cDNA for gene expression analysis.
oral examination
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