A Scintillation Proximity Assay for RNA Detection

✍ Scribed by Jianwei Liu; Patricia A. Feldman; Jonathan S. Lippy; Ekaterina Bobkova; Michael G. Kurilla; Thomas D.Y. Chung

• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 101 KB
• Volume: 289
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4944

✦ Synopsis

A homogeneous scintillation proximity assay (SPA) for detection of RNA transcripts is described. 3H-labeled RNA transcripts are hybridized in solution to biotinylated oligodeoxynucleotides (ODNs), which are then bound by streptavidin-coated, scintillant-embedded beads. Only bound 3H-labeled RNA transcripts are brought in close enough proximity to stimulate light emission from the beads. The results from this novel homogeneous assay correlated well with those obtained using the traditional filter-binding methods to measure RNA polymerase activity. The assay has been miniaturized to a 384-well format compatible with automated high-throughput screening. This SPA method has also been successfully used to probe RNA-accessible sites to hybridization, and thus should provide a useful tool for selecting effective antisense ODNs in antisense research.