Ribonucleoside Vanadyl Complex (RVC) is a potent inhibitor of various ribonucleases (1,2,3). This complex is compatible with cell fractionation methods as well as sucrose-gradient centrifugations. RVC is prepared from a modification of procedures by Lienhard using all four ribonucleosides (4).
The 200 mM stock solution is reconstituted to a green-black clear solution by incubating the sealed vial at 65°C. Once completely thawed, the entire sample should be aliquoted into smaller samples, in vials with minimal headspace, tightly sealed, and immediately frozen. The ribonucleoside vanadyl complex should be added to all buffers at a final concentration of 10 mM for maximum ribonuclease protection. The buffers should not contain EDTA since one equivalent will dissociate the complex.
We do not recommend the use of the vanadyl complex in cell free translation systems and with reverse transcriptase (5). The vanadyl complex can be used in the selective degradation of DNA while preserving RNA since pancreatic deoxyribonuclease I is not inhibited (5). More recently RVC has been shown to have use as a tissue preservative (6).
Removal of the ribonucleoside vanadyl complex from the RNA can be accomplished by adding 10 equivalents of EDTA before ethanol precipitation.
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