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RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings.

Bio-protocol (2021-03-05)
Tianran Jia, Brandon H Le
RESUMEN

Steady-state mRNA levels are determined by both the rates of transcription and degradation. Regulation of mRNA stability and/or degradation are key factors that can significantly affect mRNA levels and its biological functions. mRNA stability can be measured indirectly after transcription inhibition. This protocol described a rapid and sensitive method of mRNA stability measurement through quantitative reverse transcription PCR (RT-qPCR) after inhibition of RNA transcription by cordycepin in Arabidopsis seedlings.

MATERIALES
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Sigma-Aldrich
Medio basal Murashige y Skoog, powder, suitable for plant cell culture
Sigma-Aldrich
TWEEN® 20, for molecular biology, viscous liquid
Sigma-Aldrich
Cordycepin, from Cordyceps militaris