METHOD FOR THE SIMULTANEOUS DETECTION OF POLYGENES

摘要

A method for the simultaneous detection of polygenes involving the following steps (a) to (f): (a) the step of converting an mRNA into a cDNA by using primers for reverse transcription having the following characteristics (1) to (3): (1) having in the 3'-terminal side of the oligo dT an anchoring part represented by 5'-M(N)q-3' (wherein M represents a base selected from among A, G and C; N represents a base selected from among A, G, C and T; and q is 0 or 1); (2) having in the 5'-terminal side of the oligo dT a site for forming a solid phase; and (3) having an enzyme cleavage site between the 5'-terminus of the oligo dT and the site for forming a solid phase; (b) the step of cleaving the cDNA with a restriction enzyme; (c) the step of labeling the restriction enzyme cleavage site of the cDNA; (d) the step of forming a solid phase of the labeled cDNA fragment and recovering the same; (e) the step of cleaving the enzyme cleavage site in the 5'-terminus of the reverse transcription primer with an enzyme to thereby give a solubilized and labeled cDNA fragment; and (f) the step of subjecting the labeled cDNA fragment to two-dimensional gel electrophoresis.