INSITU DETECTION OF HUMAN BETA-1-INTERFERON MESSENGER-RNA WITH A CHEMICALLY BIOTINYLATED PROBE

This paper describes a rapid, nonradioactive in situ mRNA hybridizatio n method through the use of a chemically biotinylated DNA probe for th e detection of human beta1-IFN mRNA. The chemically biotinylated DNA p robe is characterized with respects to: (1) the percentage of modifica tion in nucleic acids analyzed by HPLC, (2) the sensitivity of detecti on, determined by reacting a series of quantities of biotinylated DNA with streptavidin-alkaline phosphatase, and (3) the hybridization spec ificity determined by hybridizing the biotinylated DNA to the in vitro synthesized beta1-IFN sense RNA dotted on the filter. The optimal con ditions of in situ mRNA hybridization were determined by investigating its specificity to the beta1-IFN mRNA in the uninduced, or RNase-trea ted induced, or induced cells.