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RNase protection assay and troubleshooting
protocol
The ribonuclease protection assay (RPA) is a highly sensitive and specific method for the detection of mRNA species. The assay was made possible by the discovery and characterization of DNA-dependant RNA polymerases from the bacteriophages SP6, T7 and T3, and ...
RNase Protection Assay
protocol
Protocol of RNase protection assay for quantitating RNA levels from either polyA+ selected RNA or crude RNA preps. (Mike A. Dyer, Harvard University) ...
RNase Protection Assay
protocol
This method can be used to detect and quantitate mRNA, to map mRNA termini and to determine the position of introns within the corresponding gene. (Gerard R. Lazo, USDA) ...
RNAse Protection Assay
protocol
A RNase protection assay protocol from J. Kennell at University of Michigan. ...
RNase Protection Assay
protocol
Protocol from the Knight lab at Loyola University.
Basic reference: Ausubel,F.M. et al. 1989.Current Protocols in Molecular Biology. John Wiley and sons, N.Y.
Templates are cloned into pGEM vectors (Promega) that contain the bacterial RNA polymerase pro ...
RNase Protection Assay System: A Versatile Technique for the Analysis of RNA
review
A promega technote written by Steven Ekenberg and Geoff Hudson.
The Promega RNase Protection Assay System provides the reagents necessary to map RNAs moreaccurately than obtained with Northern blot procedures. In addition, the system uses RNase ONE(TM) whi ...
Linearity of RNase Protection Assays
review
A technical bulletin article from Ambion on linearity of RNase protection assay. (Ambion).
The amount of protected probe is directly proportional to the amount of target mRNA in the sample. If an internal control probe or synthetic sense strand is included ...
Choice of Ribonucleases for a Ribonuclease Protection Assay
review
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 ribo ...
The Ribonuclease Protection Assay: A Powerful Tool for the Veterinary Pathologist
review
A open access journal mini review on ribonuclease protection assay. (J. B. Rottman, Millennium Pharmaceuticals)
The ribonuclease protection assay is a sensitive and accurate method to measure mRNA expression. The major advantages of the assay are that multi ...
More RNase protection assay protocols
(Curr Protoc Mol Biol. 2001 May;Chapter 4:Unit4.7.)
Gilman M.
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
Sequence-specific hybridization probes of high specific activity are prepared by cloning the probe sequence downstream of a bacteriophage promoter. The plasmid is cleaved with a restriction enzyme, and the plasmid DNA is transcribed with bacteriophage RNA polymerase, which efficiently transcribes the cloned sequence into a discrete RNA species of known specific activity and high abundance. The RNA is purified by removal of the DNA template, protein, and the unincorporated label. Alternatively, the probe is purified by gel electrophoresis, as described in a support protocol. The probe RNA is hybridized to sample RNAs and the hybridization reactions are treated with ribonuclease to remove free probe, leaving intact fragments of probe annealed to homologous sequences in the sample RNA. These fragments are analyzed by electrophoresis on a sequencing gel and the presence of the target mRNA is revealed by the appearance of an appropriately sized fragment of the probe.
Introduction to Ribonuclease Protection Assays (RPA) and RPA procedure
Ribonuclease Protection Assay (RPA assay)
Last update: 
28-Jan-2012 07:19 am
