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Measuring RNase Activity A Real-time Kinetic Analysis


The RNaseAlert QC System, when used in conjunction with a fluorometer capable of kinetic analysis, provides a rapid and ultra-sensitive assay for determining RNase A specific activity. The RNaseAlert QC System also detects a variety of other nucleases, including RNase A, T1, RNase I, micrococcal nuclease, S1 nuclease, mung bean nuclease, and Benzonase. In addition to detecting nuclease activity, this kit is useful for standardizing activities of these enzymes.


Sources of RNase

Ribonucleases (RNases) are ubiquitous in the environment, and they are present in relatively high concentrations in many biological materials. For example, the pancreas is rich in RNase (>1 mg RNase/1 g tissue) and is the source for most commercially produced RNase A (and its glycosylated derivative, RNase B).

RNase A is an endonuclease that cleaves 3' of C and U residues. It is commonly used in molecular biology applications such as the removal of contaminating RNA from DNA preparations and ribonuclease protection assays (RPA). Ambion sells an RPA grade RNase A (Cat #2272) that is equivalent to molecular biology grade RNase A from other suppliers, as well as affinity-purified RNase A (Cat #2270 and #2271), which is recommended when the absence of DNase and other nonspecific nuclease activities is essential.


Measuring Specific Activity


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