Choice of Ribonucleases for a Ribonuclease Protection Assay

A technical bulletin article on Choice of Ribonucleases for a Ribonuclease Protection Assay. (Ambion inc).

Ribonuclease protection assays (RPAs) are a technique used for detection and quantitation of specific RNAs. The method is based on the ability of ribonuclease to specifically degrade single-stranded RNA while leaving intact a labeled antisense RNA probe that is hybridized to its target. Ribonuclease protection assays are more sensitive than Northern blots for the detection of mRNAs, more tolerant of partially degraded RNA, and are able to distinguish between transcripts of multi-gene families that co-migrate on Northern blots. Using a high specific-activity probe (approximately 109 cpm/µg), a moderately abundant mRNA such as ß-actin can easily be detected in sub-microgram quantities of total RNA.

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