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Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR

mRNA decay is an important process in post-transcriptional regulation; in addition, it plays a crucial role in plant development and response to stress. The development of new tools to quantify mRNA decay intermediates is thus important to better characterize the dynamic of mRNA decay in various con...

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Autores principales: Boubegtitene, Alexandre, Merret, Rémy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9571659/
https://www.ncbi.nlm.nih.gov/pubmed/36235485
http://dx.doi.org/10.3390/plants11192616
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author Boubegtitene, Alexandre
Merret, Rémy
author_facet Boubegtitene, Alexandre
Merret, Rémy
author_sort Boubegtitene, Alexandre
collection PubMed
description mRNA decay is an important process in post-transcriptional regulation; in addition, it plays a crucial role in plant development and response to stress. The development of new tools to quantify mRNA decay intermediates is thus important to better characterize the dynamic of mRNA decay in various conditions. Here, we applied droplet digital PCR (ddPCR), a recent and precise PCR technology, to determine mRNA half-life in Arabidopsis seedlings. We demonstrated that ddPCR can correctly assess mRNA half-life from a wide variety of transcripts in a reproducible manner. We also demonstrated that thanks to multiplexing mRNA, the half-life of multiple transcripts can be followed in the same reaction. As ddPCR allows precise quantification, we proposed that this approach is highly suitable when a low amount of RNA is available; for the detection of many targets or for the analysis of lowly expressed transcripts.
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spelling pubmed-95716592022-10-17 Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR Boubegtitene, Alexandre Merret, Rémy Plants (Basel) Technical Note mRNA decay is an important process in post-transcriptional regulation; in addition, it plays a crucial role in plant development and response to stress. The development of new tools to quantify mRNA decay intermediates is thus important to better characterize the dynamic of mRNA decay in various conditions. Here, we applied droplet digital PCR (ddPCR), a recent and precise PCR technology, to determine mRNA half-life in Arabidopsis seedlings. We demonstrated that ddPCR can correctly assess mRNA half-life from a wide variety of transcripts in a reproducible manner. We also demonstrated that thanks to multiplexing mRNA, the half-life of multiple transcripts can be followed in the same reaction. As ddPCR allows precise quantification, we proposed that this approach is highly suitable when a low amount of RNA is available; for the detection of many targets or for the analysis of lowly expressed transcripts. MDPI 2022-10-05 /pmc/articles/PMC9571659/ /pubmed/36235485 http://dx.doi.org/10.3390/plants11192616 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Technical Note
Boubegtitene, Alexandre
Merret, Rémy
Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR
title Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR
title_full Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR
title_fullStr Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR
title_full_unstemmed Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR
title_short Monitoring mRNA Half-Life in Arabidopsis Using Droplet Digital PCR
title_sort monitoring mrna half-life in arabidopsis using droplet digital pcr
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9571659/
https://www.ncbi.nlm.nih.gov/pubmed/36235485
http://dx.doi.org/10.3390/plants11192616
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