METHOD OF DETECTION OF DIFFERENTIALLY EXPRESSING TEMPLATE RNAS AND CLONING THE CORRESPONDING CDNA FRAGMENTS Russian patent published in 1998 - IPC

FIELD: biotechnology, genetic engineering. SUBSTANCE: for detection of differentially expressing mRNAs (demRNAs) mRNAs are isolated from comparable cell types and the corresponding cDNAs are synthesized. cDNAs are treated with restriction endonucleases producing many small fragments. From each separate set of restriction fragments the corresponding fragments with either only 3'-terminal or only 5'-terminal regions of mRNAs of each cell type are selected. Obtained sets are labeled and subdivided their obtaining noncrossing subsets of cDNA-fragments which are analyzed by gel-electrophoresis. Pictures of their separation are compared and cDNA-fragments with different signal intensity are detected. For cloning cDNA-fragments, corresponding demRNAs are isolated from gel, amplificated by polymerase chain reaction and inserted in vector suitable for cloning. Invention can be used in medicine for disease diagnosis and scientific investigations for etiology study at genetic level. EFFECT: improved method of detection. 16 cl, 6 dwg