A nonradioactive assay for poly(a)-specific ribonuclease activity by methylene blue colorimetry.
A simple nonradioactive assay, which was based on the specific shift of the absorbance maximum of methylene blue induced by its intercalation into poly(A) molecules, was developed for poly(A)-specific ribonuclease (PARN). A good linear relationship was found between the absorbance at 662 nm and the poly(A) concentration. The assay conditions, including the concentration of methylene blue, the incubation temperature and time, and the poly(A) concentration were evaluated and optimized.
Cheng
Yuan
Y
Life Science Division, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China.
Liu
Wei-Feng
WF
Yan
Yong-Bin
YB
Zhou
Hai-Meng
HM
D003124:Colorimetry / Q000379:methods*; D003947:Diagnostic Techniques, Radioisotope; D005095:Exoribonucleases / Q000378:metabolism*; D006801:Humans; D008751:Methylene Blue / Q000032:analysis*; D011061:Poly A / Q000378:metabolism*; D013057:Spectrum Analysis; D013379:Substrate Specificity; D013696:Temperature
D005095:Exoribonucleases; D008751:Methylene Blue; D011061:Poly A