Vanadyl-ribonucleoside complex

Talib, S., and J.E. Hearst. 1983. Initiation of RNA synthesis in vitro by vesicular stomatitis virus Single internal initiation in the presence of aurintricarboxylic acid and vanadyl ribonucleoside complexes. Nucleic Acids Res 11 7031. 

The quick blot procedure, used for mRNA (not efficient for rRNA or tRNA), is based on the same principle (Costanzi and Gillespie, 1987). A cell pellet is resuspended in Hanks balanced salt solution (Gibco), containing 50 pg/ml of cycloheximide and 10 mM vanadyl-ribonucleoside complexes, to about 10 -10 cells/ml. The cells are incubated for 10 min at 37 C after adding 1 vol. of proteinase K stock solution. Then, 0.05 vol. of 10% Brij 35 is added and mixed, followed by mixing with 0.05 vol. of sodium deoxycholate. The subsequent steps with fresh hot Nal are as for DNA, except that 5 M Nal containing 0.5% Brij is used for dilution. 

Vanadyl-ribonucleoside complex method for preparation of nuclear and cytoplasmic RNA... 

Prepare the DNAase I mix, sufficient for four reactions. 20 pL of DNAase I digestion buffer, 1 pL of vanadyl ribonucleoside complex, I pL of RNAase-free DNAase I (10 U/pL) Let stand at room temperature for 5 min. 

Berger, S.L., and C.S. Birkenmeier. 1979. Inhibition of intractable nucleases with ribonucleoside-vanadyl complexes Isolation of messenger ribonucleic acid from resting lymphocytes. Biochemistry 18 5143. 

Berger, S.L. (1987). Isolation of cytoplasmic RNA ribonucleoside-vanadyl complexes. Meth. Enzymol. 152, 227-234. 

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