Isolation of nuclei in tagged cell types (intact), rna extraction and ribosomal rna degradation to prepare material for rna-seq
2018
Reynoso, Mauricio A. | Pauluzzi, Germain C. | Cabanlit, Sean | Velasco, Joel | Bazin, Jérémie | Deal, Roger | Brady, Siobhan | Sinha, Neelima | Bailey-Serres, Julia | Kajala, Kaisa | Center for Plant Cell Biology - Department of Botany and Plant Sciences ; University of California [Riverside] (UC Riverside) ; University of California (UC)-University of California (UC) | Institut des Sciences des Plantes de Paris-Saclay (IPS2 (UMR_9213 / UMR_1403)) ; Institut National de la Recherche Agronomique (INRA)-Université Paris-Sud - Paris 11 (UP11)-Université Paris Diderot - Paris 7 (UPD7)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS) | Emory University [Atlanta, GA] | University of California [Davis] (UC Davis) ; University of California (UC) | Department of Plant Biology ; University of California [Davis] (UC Davis) ; University of California (UC)-University of California (UC) | Universiteit Utrecht / Utrecht University [Utrecht]
International audience
Mostrar más [+] Menos [-]Inglés. Gene expression is dynamically regulated on many levels, including chromatin accessibility and transcription. In order to study these nuclear regulatory events, we describe our method to purify nuclei with Isolation of Nuclei in TAgged Cell Types (INTACT). As nuclear RNA is low in polyadenylated transcripts and conventional pulldown methods would not capture non-polyadenylated pre-mRNA, we also present our method to remove ribosomal RNA from the total nuclear RNA in preparation for nuclear RNA-Seq.
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