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A homogeneous method to quantify mRNA levels: a hybridization of RNase protection and scintillation proximity assay
M K Kenrick1, L Jiang, C L Potts
1Amersham International plc, Cardiff Laboratories, Forest Farm, Whitchurch, Cardiff CF4 7YT, UK.
Abstract:
A novel method to measure mRNA levels has been developed by combining the detection capabilities of RNase protection (RPA) with the quantification advantages of scintillation proximity assay (SPA) technology. Sample processing is reduced to the addition of a single reagent post RNase digestion. As a model system, the inducible expression of rat apolipoprotein-A1 mRNA has been measured by both traditional gel-based RPAs and the SPA-based RPA assay. Results demonstrate that the ribonuclease protection proximity assay (RiPPA) faithfully reproduces the gel-based results and is at least as sensitive as many existing methods.