Biomolecular Technology - Modulo 2 - Laboratorio (2008/2009)

Learning outcomes

Southern-blot (based on hybridization between nucleic acids) and RT-PCR (based on PCR) performed on transgenic and wild-type plants, will allow the student to apply the knowledges acquired.

1) SOUTHERN BLOT ANALYSIS OF TRANSGENIC PLANTS: GENOMIC DNA EXTRACTION, QUANTIFICATION, RESTRICTION DIGESTION, ELECTROPHORESIS OD DIGESTED DNA, CAPILLARY BLOTTING, FIXING THE DNA TO THE MEBRANE, LABELING NONRADIOACTIVE PROBE, PREHYBRIDIZATION, HYBRIDIZATION, WASHING, DETECTION OF HYBRIDS BY CHEMIOLUMINESCENT SUBSTRATE

2) RT-PCR: RNA EXTRACTION, QUANTIFICATION, DNase TREATMENT, SYNTHESIS OF cDNA, PCR

Delivery of a detailed report regarding the experiments performed in the laboratory.