A scintillation proximity assay for RNA detection

被引：11

作者：

Liu, JW ^{[1
]}

Feldman, PA ^{[1
]}

Lippy, JS ^{[1
]}

Bobkova, E ^{[1
]}

Kurilla, MG ^{[1
]}

Chung, TDY ^{[1
]}

机构：

[1] Dupont Co, Expt Stn, Wilmington, DE 19880 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2001年 / 289卷 / 02期

关键词：

scintillation proximity assay; RNA detection; RNA accessibility; RNA polymerase; in vitro transcription;

D O I：

10.1006/abio.2000.4944

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A homogeneous scintillation proximity assay (SPA) for detection of RNA transcripts is described. H-3-labeled RNA transcripts are hybridized in solution to biotinylated oligodeoxynucleotides (ODNs), which are then bound by streptavidin-coated, scintillant-embedded beads. Only bound H-3-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. (C) 2001 Academic Press.

引用

页码：239 / 245

页数：7

共 12 条

[1] The effect of structure in a long target RNA on ribozyme cleavage efficiency [J].