Determination of gene expression by specific capture of RNA and direct quantification thereof by free-zone capillary electrophoresis

摘要

Determination of gene expression by specific capture of RNA and direct quantification thereof by free-zone capillary electrophoresis. Method for determination of gene expression by direct detection of transcripts by means of free-zone capillary electrophoresis for biological samples. Purified or semi-purified RNA samples are hybridized with an oligonucleotide or previously selected specific probe, labelled with biotin and belonging to a gene coding sequence. Magnetic spheres loaded with streptavidin are added to the hybridization solution containing the specific hybrid. A magnetic system is then applied which selects the streptavidin- biotin-oligonucleotide or specific RNA probe complex in the wall of the tube, the complex is washed with astringent solutions and the specific RNA is then eluted. The RNA obtained is injected into a free-zone capillary electrophoresis system and the signal obtained by the detector allows direct quantification of the sample.