An assay to determine the kinetics of RNA cleavage.
To evaluate some synthetic catalysts that mimic ribonuclease, a quantitative assay has been developed that measures the number of phosphate diester bonds cleaved in a polymeric RNA substrate. This assay involves determining the number of 5'-oligonucleotide termini produced during the cleavage, using polyuridylic acid as the substrate. Samples withdrawn from the kinetic run are treated with venom exonuclease (phosphodiesterase I), and the increase in the concentration of uridine is determined by high-performance liquid chromatography. A related assay has been developed to monitor the catalyzed cleavage of the dinucleotide uridylyl(3'----5') uridine (UpU).
Corcoran
R
R
Labelle
M
M
Czarnik
A W
AW
Breslow
R
R
D002851:Chromatography, High Pressure Liquid; D015226:Dinucleoside Phosphates; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D011072:Poly U / Q000378:metabolism; D012313:RNA / Q000032:analysis*; D012260:Ribonucleases / Q000378:metabolism; D014529:Uridine / Q000031:analogs & derivatives / Q000032:analysis; D014542:Uridine Monophosphate / Q000031:analogs & derivatives / Q000032:analysis
D015226:Dinucleoside Phosphates; C026519:uridine monophosphate-uridine; D011072:Poly U; D012313:RNA; D014542:Uridine Monophosphate; D012260:Ribonucleases; D014529:Uridine