Fast Procedure with Sensitive Results
Figure 1. Schematic of RNaseAlert™ Procedure.
The most common method for detecting RNase contamination in solutions is to incubate an RNA substrate with a test solution and then check for degradation of the RNA by ethidium bromide stained agarose gel electrophoresis. This assay has very low sensitivity and requires a subjective judgment. Other more sensitive tests for RNase require a radio-labeled RNA substrate or a fluorescence polarization instrument and are time consuming and costly. The RNaseAlert technology is extremely sensitive and can detect 0.5 pg RNase A or less, much less than most common RNase tests. In addition, the RNaseAlert procedure is neither time-consuming nor requires specialized equipment. The assay is non-toxic and includes all the necessary reagents to perform your own quality control assays. Ambion scientists have successfully used the RNaseAlert kit to detect many types of RNases from various sources. While stable in the absence of RNase, the fluorescence substrate can be cleaved under a wide range of conditions, making this assay useful for examining most molecular biology solutions or surfaces for contaminating RNases.
Detect RNases in Any Lab
The RNaseAlert Lab Test Kit is designed to indicate contaminating RNases in almost any solution by a clear visual read-out. The lyophilized substrate is dissolved in the provided reaction buffer, the sample to be tested is added, and the assay is incubated at 37°C. The results are visualized by irradiating the tube with a UV light source (transilluminator or hand held light) and looking for a bright green fluorescent glow. The fluorescence intensity will be directly proportional to the amount of RNase contamination. This assay is typically completed in less than one hour. For a permanent record of the results, tubes can be photographed using a gel documentation system. If a quantitative value is needed, the fluorescence can be quantified in a fluorometer. For routine or higher throughput RNase monitoring, the RNaseAlert QC System is recommended.
High Throughput RNase Detection
Figure 2. Real-time Fluorescent Monitoring of RNase A Activity. The indicated amounts of RNase A (0-0.2 pg) were monitored in the RNaseAlert QC System during a 1 hour incubation at 37°C.