RNaseAlert™ Lab Test Kit v2 (Invitrogen™)

RNaseAlert™ Lab Test Kit v2 detects RNase activity in a convenient and sensitive fluorimetric assay that delivers results in real time. This kit is suitable for testing small numbers of sample and ensuring that solutions, tubes, tips, etc. in your lab are RNase-free. This new version contains a more sensitive substrate that is ∼60% more sensitive than the previous version. The kit contains sufficient reagents for 25 reactions.

• Detects as little as ∼0.3 pg of RNase A with a fluorometer
• Simple, straight-forward 30-minute assay

The RNaseAlert™ Lab Test Kit v2 uses a new, more sensitive, novel RNA substrate tagged with a fluorescent reporter molecule (fluor) on one end and a quencher on the other. In the absence of RNases, the physical proximity of the quencher dampens fluorescence from the fluor to extremely low levels. When RNases are present, however, the RNA substrate is cleaved, and the fluor and quencher are spatially separated in solution. This causes the fluor to emit a bright green signal when excited by light of the appropriate wavelength.

Fluorescence can be readily detected with a filter-based or monochromator-based fluorometer. Since the fluorescence of the RNaseAlert™ substrate increases over time when RNase activity is present, results monitored with a fluorometer can be evaluated kinetically. The sequence of the RNaseAlert™ substrate has been carefully optimized to detect several RNases, including RNase A, RNase T1, RNase I, micrococcal nuclease, S1 nuclease, mung bean nuclease, and Benzonase™.

Rapid and Convenient Protocol
The RNaseAlert™ procedure requires just minutes to set up. The lyophilized, fluorescent substrate is reconstituted in the supplied tube and incubated with the test solution for a few minutes. The results are then read on a fluorometer. Samples that noticeably fluoresce compared to the negative control are contaminated and should not be used with RNA. Nuclease-free water and RNase A are provided for use as controls.