This paper describes a rapid, nonradioactive in situ mRNA hybridization method through the use of a chemically biotinylated DNA probe for the detection of human β1-IFN mRNA. The chemically biotinylated DNA probe is characterized with respects to: (1) the percentage of modification in nucleic acids analyzed by HPLC, (2) the sensitivity of detection, determined by reacting a series of quantities of biotinylated DNA with streptavidin-alkaline phosphatase, and (3) the hybridization specificity determined by hybridizing the biotinylated DNA to the in vitro synthesized β1-IFN sense RNA dotted on the filter. The optimal conditions of in situ mRNA hybridization were determined by investigating its specificity to the β1-IFN mRNA in the uninduced, or RNase-treated induced, or induced cells.
ASJC Scopus subject areas
- Pathology and Forensic Medicine
- Cell Biology