A METHOD FOR SIMULTANEOUS DETECTION OF MULTIPLE MESSENGER-RNAS USING DIGOXIGENIN AND RADIOISOTOPIC CRNA PROBES

We present a simple, reliable method for simultaneous detection of two distinct mRNAs within the same tissue sample by double in situ hybrid ization histochemistry. Sections are hybridized with a cocktail of rad iolabeled and digoxigenin-labeled cRNA probes. The digoxigenin-labeled probe is detected with an alkaline phosphatase (AP)-dependent chromog en reaction and then the radiolabeled probe is detected by conventiona l autoradiography. The sensitivities of the two detection methods are comparable and demonstrate the feasibility of using double in situ hyb ridization histochemistry to investigate the co-localization and co-re gulation of mRNAs that are expressed at relatively low levels.