Core Facilities

BIACORE

Clinical Immunology
Laboratory

Myositis Testing

DNA Sequencing

Flow Cytometry

Imaging Core Facility

Microarray Research
Facility

RPA/RNA/In Situ
Hybridization Research
Core

Mouse Genome
Manipulation Facility
(Transgenics)

Signal Transduction
Core Facility

Small Animal MRI

Shared Facilities

Laboratory Animal
Resource Center (LARC)

Institutional Animal
Care and Use
Committee (IACUC)

RIBONUCLEASE PROTECTION ASSAY (RPA) is a method to simultaneously quantify multiple mRNAs in a total RNA extract.  This method utilized 32P-labeled RNA probes that hybridize to their cognate mRNA creating a region of double-stranded RNA that is resistant to RNase digestion.  These dsRNAs are resolved on a sequencing gel and visualized and quantitated by phosphorimager.  The user can select from a range of multiple-probe template sets (cytokines, apotosis genes, stress response genes, DNA repair genes etc.) available from PharMingen (www.pharmingen.com).

RPA phospharimage using cytokine probes performed on RNA extracted from the spinal cords of wild-type and transgenic mice overexpressing a mutant form (G93A) of human superoxide dismutase 1.

The overall procedure includes:

1. Total RNA extraction from the sample

2. Radio-labeling of the probe set

3. Hybridization of probes with RNA followed by RNase digestion.

4. Gell electrophoresis of the hybridized and RNase-digested mRNA and visualization/quantitation by phosphorimages

Charges:

RNA preparation and RPA analysis (1 gel - 20 samples) for OMRF investigators - $200
RNA preparation and RPA analysis (1 gel - 20 samples for OUHSC investigators - $400
(These charges do not include the cost of probe sets or 32P radioisotope)

Please contact Dr. Charles Stewart, 271-7583 for details.