基于 Tn5 转座酶纯化技术的 ATAC-Seq 的表观遗传学应用。
Epigenetic Application of ATAC-Seq Based on Tn5 Transposase Purification Technology.
机构信息
Department of Critical Care Medicine, Shunde Hospital Affiliated of Jinan University, 528305 Foshan, China.
Meng Yi Center Limited, Macau 999078, China.
出版信息
Genet Res (Camb). 2022 Aug 11;2022:8429207. doi: 10.1155/2022/8429207. eCollection 2022.
BACKGROUND
Assays of transposase accessible chromatin sequencing (ATAC-seq) is an efficient assay to investigate chromatin accessibility, which depends on the activity of a robust Tn5 transposase to fragment the genome while cutting in the sequencing adapters.
METHODS
We propose reliable approaches for purifying hyperactive Tn5 transposase by chitin magnetic bead sorting. Double-stranded DNA of J76 cells and 293T cells were digested and subjected to tagmentation as test samples with Tn5 transposase, and libraries were established and sequenced. Sequencing data was then analyzed for peak calling, GO enrichment, and motif analysis.
RESULTS
We report a set of rapid, efficient, and low-cost methods for ATAC-seq library construction and data analysis, through large-scale and rapid sequencing. These methods can provide a reference for the study of epigenetic regulation of gene expression.
背景
转座酶可及染色质测序（ATAC-seq）分析是一种高效的研究染色质可及性的方法，该方法依赖于活性很强的 Tn5 转座酶来在测序接头处切割的同时将基因组片段化。
方法
我们提出了通过几丁质磁珠分选来纯化高活性 Tn5 转座酶的可靠方法。用 Tn5 转座酶对 J76 细胞和 293T 细胞的双链 DNA 进行消化和标签酶切，构建文库并进行测序。然后对测序数据进行峰调用、GO 富集和基序分析。
结果
我们通过大规模快速测序，报告了一套快速、高效、低成本的 ATAC-seq 文库构建和数据分析方法。这些方法可以为研究基因表达的表观遗传调控提供参考。
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