Creative Biolabs is a well-recognized expert in the field of RNA assay. With over ten years of extensive experience and advanced platforms, our scientists are confident in offering the best-fit nuclease protection assay to assist your valuable projects in a high-quality manner.
Nuclease protection assay is extremely sensitive for the detection, quantitation, and mapping of specific RNAs in complex mixtures of total cellular RNA. Nuclease protection assays include both S1 nuclease assay and RNase protection assay, which differ only in the probe used (i.e., RNA and DNA probes, respectively). The nuclease protection assay is a nucleic acid hybridization-based technique that can be paired with other methods to improve specificity and has the potential to be developed into a point-of-need device. This method relies on solution-phase hybridization between the target mRNA and radiolabeled complementary RNA molecules. Moreover, the technique can simultaneously identify multiple RNA molecules even at low total concentrations. Nuclease protection assay is a powerful method to identify target RNA in the RNA sample extracted from cells or tissue extracts, primarily because of its high sensitivity.
For this nuclease protection assay, radioactively labeled DNA or RNA probes are synthesized that are complementary to the target RNA to be analyzed. The probe is then mixed with sample RNA incubated to permit the hybridization. Following the hybridization step, an S1 nuclease or RNase is used to digest away the remaining unhybridized and unprotected probe and RNA sample. The nucleases are then inactivated, and the protected probe fragments are precipitated from the reaction mixture and analyzed on a high-resolution, denaturing polyacrylamide gel. The signal of radioactively labeled probes can then be visualized and detected by autoradiography or other methods.
Nuclease protection assay has been used in quantitation of RNA levels, RNA mapping, determining gene transcription levels, and even determining sites and extents of protein-RNA interactions for decades. Due to the high resolution of the polyacrylamide gel system used to analyze the protected probe, nuclease protection assay is well suited for mapping positions of external and internal junctions in RNA-such as transcription initiation and termination sites and intron/exon boundaries. Moreover, nuclease protection assay can be used to discriminate between closely related targets, for example, members of multigene families, by designing the probes to span the regions where the related genes differ the most.
As a pioneer and undisputed global leader in the field of mRNA-based assays, Creative Biolabs is professional in applying advanced nuclease protection strategies to satisfy various project demands. Based on our extensive experience in nuclease protection assay, we have won a great reputation among our worldwide clients for successfully detecting RNA. Please feel free to contact us for more details.