RNaseAlert™ QC System v2 (Invitrogen™)

The RNaseAlert™ QC System v2 detects RNase activity in a convenient and sensitive fluorimetric assay that delivers results in real time. This version contains a substrate that is ∼60% more sensitive than the previous version. The kit contains sufficient reagents for 5 x 96 (480) high-throughput assays.

Features of RnaseAlert™ QC System v2:

• Detects as little as ∼0.3 pg of RNase A with a fluorometer
• Simple, straight-forward, 30-minute assay
• Designed to monitor RNase activity in column fractions during protein purification
• Use with DNaseAlert™ QC System to quantitatively detect both RNases and DNases in the same sample

The RNaseAlert™ QC System 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.

Accessory Product
The RNaseAlert™ QC System v2 is fully compatible with the DNaseAlert™ QC System. Since the DNaseAlert™ substrate contains a fluorescent tag that is spectrally distinct from RNaseAlert™ substrate, the two kits can be used simultaneously for quantitative real-time analyses of both RNases and DNases within a single sample.