Unit: Molecular Biology I
Bachelor degree in Molecular Biology, II year
A detailed description of the molecular biology of the prokaryotic gene.
The practicals are an intense "primer" on molecular biology techniques. You'll prepare competent E. coli cells and transform them with genomic DNA (lambda phage) digested with two restriction enzymes (EcoRI and BamHI). After plating you'll screen the colonies using the blue-white method (beta-galactosidase and X-Gal). Then you'll perform a mini preparation of plasmidic DNA from selected colonies, and digest them to retrieve the cloned fragment.
Another experiment will be the extraction of genomic DNA from your saliva, that will be used as template for a diagnostic PCR that amplify less than 100bp of the CTFR gene. A mutated allele of the gene, laking 3 bases, is associated with a disease. Using a polyacrylamide gel we will be able to display which alleles our genome carries.
Please use 2010-2011 Course website as a reference.