Quantification of all 12 canonical ribonucleotides by real-time fluorogenic in vitro transcription.

ABSTRACT: Enzymatic methods to quantify deoxyribonucleoside triphosphates have existed for decades. In contrast, no general enzymatic method to quantify ribonucleoside triphosphates (rNTPs), which drive almost all cellular processes and serve as precursors of RNA, exists to date. ATP can be measured with an enzymatic luminometric method employing firefly luciferase, but the quantification of other ribonucleoside mono-, di-, and triphosphates is still a challenge for a non-specialized laboratory and practically impossible without chromatography equipment. To allow feasible quantification of ribonucleoside phosphates in any laboratory with typical molecular biology and biochemistry tools, we developed a robust microplate assay based on real-time detection of the Broccoli RNA aptamer during in vitro tr

SUBMITTER: Purhonen J 

PROVIDER: S-EPMC10783517 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature