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低输入 ATAC&mRNA-seq 方案用于同时分析染色质可及性和基因表达。

Low-input ATAC&mRNA-seq protocol for simultaneous profiling of chromatin accessibility and gene expression.

机构信息

Epigenetics Innovation Lab, Center for Epigenetics Research, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.

Integrative Bioinformatics Support Group, Epigenetics and Stem Cell Biology Laboratory, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.

出版信息

STAR Protoc. 2021 Aug 27;2(3):100764. doi: 10.1016/j.xpro.2021.100764. eCollection 2021 Sep 17.

Abstract

We present a simple, fast, and robust protocol (low-input ATAC&mRNA-seq) to simultaneously generate ATAC-seq and mRNA-seq libraries from the same cells in limited cell numbers by coupling a simplified ATAC procedure using whole cells with a novel mRNA-seq approach that features a seamless on-bead process including direct mRNA isolation from the cell lysate, solid-phase cDNA synthesis, and direct tagmentation of mRNA/cDNA hybrids for library preparation. It enables dual-omics profiling from limited material when joint epigenome and transcriptome analyses are needed. For complete details on the use and execution of this protocol, please refer to Li et al. (2021).

摘要

我们提出了一种简单、快速、稳健的方案(低输入 ATAC&mRNA-seq),通过将简化的全细胞 ATAC 方案与一种新型的 mRNA-seq 方法相结合,在有限的细胞数量中从同一细胞中同时生成 ATAC-seq 和 mRNA-seq 文库,该方法具有无缝的珠上过程,包括直接从细胞裂解物中分离 mRNA、固相 cDNA 合成以及直接对 mRNA/cDNA 杂交物进行片段化以进行文库制备。当需要联合进行表观基因组和转录组分析时,它可以从有限的材料中进行双重组学分析。有关此方案的使用和执行的详细信息,请参阅 Li 等人(2021 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7c04/8403761/9c8db105b9f0/fx1.jpg

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