Biomolecular Technology (2009/2010)

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Course code
4S00175
Credits
8
Coordinator
Massimo Delledonne
Academic sector
AGR/07 - AGRICULTURAL GENETICS BIO/11 - MOLECULAR BIOLOGY
Language of instruction
Italian
Teaching is organised as follows:
Activity Credits Period Academic staff Timetable
Teoria 2 2nd Semester Massimo Delledonne
Laboratorio 6 2nd Semester Diana Bellin, Sara Zenoni

Lesson timetable

2nd Semester
Activity Day Time Type Place Note
Teoria Monday 2:30 PM - 5:30 PM lesson Lecture Hall C from Apr 19, 2010  to Jun 15, 2010
Teoria Friday 2:30 PM - 5:30 PM lesson Lecture Hall H from Apr 16, 2010  to Jun 15, 2010
Laboratorio Wednesday 8:30 AM - 12:30 PM lesson Laboratory Laboratorio di Biotecnologie genetiche from Apr 28, 2010  to Jun 15, 2010
Laboratorio Friday 8:30 AM - 12:30 PM lesson Laboratory Laboratorio di Biotecnologie genetiche from May 14, 2010  to Jun 15, 2010
Laboratorio Friday 1:30 PM - 5:30 PM lesson Laboratory Laboratorio di Biotecnologie genetiche from May 14, 2010  to Jun 15, 2010

Learning outcomes

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.

Syllabus

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.

Assessment methods and criteria

oral examination

Studying