在标记细胞类型中分离细胞核（INTACT）、提取RNA并降解核糖体RNA以制备用于RNA测序的材料。
Isolation of Nuclei in Tagged Cell Types (INTACT), RNA Extraction and Ribosomal RNA Degradation to Prepare Material for RNA-Seq.
作者信息
Reynoso Mauricio A, Pauluzzi Germain C, Cabanlit Sean, Velasco Joel, Bazin Jérémie, Deal Roger, Brady Siobhan, Sinha Neelima, Bailey-Serres Julia, Kajala Kaisa
机构信息
Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, CA, USA.
IPS2, Institute of Plant Science-Paris Saclay (CNRS-INRA), University of Paris-Saclay, Orsay, France.
出版信息
Bio Protoc. 2018 Apr 5;8(7):e2458. doi: 10.21769/BioProtoc.2458.
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.
基因表达在多个层面受到动态调控，包括染色质可及性和转录。为了研究这些细胞核调控事件，我们描述了一种利用标记细胞类型细胞核分离法（INTACT）纯化细胞核的方法。由于细胞核RNA中多聚腺苷酸化转录本含量较低，传统的下拉方法无法捕获非多聚腺苷酸化的前体mRNA，我们还介绍了一种从总细胞核RNA中去除核糖体RNA的方法，以用于细胞核RNA测序。
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