Yang N, Wang S, Faber J E
Department of Physiology, University of Carolina, Chapel Hill, NC.
Methods Mol Med. 1999;30:201-19. doi: 10.1385/1-59259-247-3:201.
Ribonuclease protection assay (RPA) is a sensitive solution hybridization method for quantitation of specific RNAs (1-3). The method is based on the ability of single-strand specific ribonuclease to degrade single-stranded RNA while leaving intact fragments of labeled antisense RNA probe, which are annealed to homologous sequence in the sample RNA. After ribonuclease digestion, the hybridized portion of the probe ("protected fragment") can be visualized via electrophoresis of the mixture on a denaturing polyacrylamide gel followed by autoradiography.
核糖核酸酶保护分析(RPA)是一种用于定量特定RNA的灵敏的溶液杂交方法(1-3)。该方法基于单链特异性核糖核酸酶降解单链RNA的能力,同时使标记的反义RNA探针的完整片段保持不变,这些片段与样品RNA中的同源序列退火。核糖核酸酶消化后,探针的杂交部分(“保护片段”)可通过将混合物在变性聚丙烯酰胺凝胶上进行电泳,然后进行放射自显影来可视化。