Isolation of Nuclei in Tagged Cell Types (INTACT), RNA Extraction and Ribosomal RNA Degradation to Prepare Material for RNA-Seq.

Autor: Reynoso MA; Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, CA, USA., Pauluzzi GC; Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, CA, USA., Cabanlit S; Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, CA, USA., Velasco J; Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, CA, USA., Bazin J; IPS2, Institute of Plant Science-Paris Saclay (CNRS-INRA), University of Paris-Saclay, Orsay, France., Deal R; Emory University, Atlanta, GA, USA., Brady S; Department of Plant Biology, UC Davis, Davis, CA, USA.; Genome Center, UC Davis, Davis, CA, USA., Sinha N; Department of Plant Biology, UC Davis, Davis, CA, USA., Bailey-Serres J; Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, CA, USA., Kajala K; Department of Plant Biology, UC Davis, Davis, CA, USA.; Genome Center, UC Davis, Davis, CA, USA.
Jazyk: angličtina
Zdroj: Bio-protocol [Bio Protoc] 2018 Apr 05; Vol. 8 (7), pp. e2458. Date of Electronic Publication: 2018 Apr 05 (Print Publication: 2018).
DOI: 10.21769/BioProtoc.2458
Abstrakt: 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.
(Copyright © 2018 The Authors; exclusive licensee Bio-protocol LLC.)
Databáze: MEDLINE
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