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将全基因组染色质转座酶可接近性分析(Omni-ATAC)协议优化用于冷冻保存的大鼠脂肪和肌肉组织中的染色质可及性分析。

Optimization of the Omni-ATAC protocol to chromatin accessibility profiling in snap-frozen rat adipose and muscle tissues.

作者信息

Nair Venugopalan D, Vasoya Mital, Nair Vishnu, Smith Gregory R, Pincas Hanna, Ge Yongchao, Douglas Collin M, Esser Karyn A, Sealfon Stuart C

机构信息

Department of Neurology, Center for Advanced Research on Diagnostic Assays, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.

Department of Computer Sciences, Columbia University, New York, NY 10027, USA.

出版信息

MethodsX. 2022 Apr 1;9:101681. doi: 10.1016/j.mex.2022.101681. eCollection 2022.

Abstract

ATAC-seq is a fast and sensitive method for the epigenomic profiling of open chromatin and for mapping of transcription factor binding sites [1]. Despite the development of the Omni-ATAC protocol for the profiling of chromatin accessibility in frozen tissues [2], studies in adipose tissue have been restricted due to technical challenges including the high lipid content of adipocytes and reproducibility issues between replicates. Here, we provide a modified Omni-ATAC protocol that achieves high data reproducibility in various tissue types from rat, including adipose and muscle tissues [3].•This protocol describes a methodology that enables chromatin accessibility profiling from snap-frozen rat adipose and muscle tissues.•The technique comprises an optimized bead-based tissue homogenization process that substitutes to Dounce homogenization, reduces variability in the experimental procedure, and is adaptable to various tissue types.•In comparison with the Omni-ATAC protocol, the method described here results in improved ATAC-seq data quality that complies with ENCODE quality standards.

摘要

ATAC-seq是一种用于开放染色质表观基因组分析和转录因子结合位点图谱绘制的快速且灵敏的方法[1]。尽管已经开发出用于冷冻组织中染色质可及性分析的Omni-ATAC方案[2],但由于技术挑战,包括脂肪细胞的高脂含量以及重复样本之间的可重复性问题,脂肪组织的研究受到了限制。在此,我们提供了一种改良的Omni-ATAC方案,该方案在大鼠的各种组织类型(包括脂肪和肌肉组织)中实现了高数据可重复性[3]。

•本方案描述了一种能够对速冻大鼠脂肪和肌肉组织进行染色质可及性分析的方法。

•该技术包括一个基于珠子的优化组织匀浆过程,该过程替代了Dounce匀浆法,减少了实验过程中的变异性,并且适用于各种组织类型。

•与Omni-ATAC方案相比,本文所述方法可提高ATAC-seq数据质量,符合ENCODE质量标准。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f1d6/9027329/f193648614c5/ga1.jpg

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