ATAC-seq(Assay for Transposase-Accessible Chromatin with high throughput sequencing)染色质可及性分析是表观领域也是一种非常重要的测序方法,常见的万金油套路有两种:1.ATAC-seq+RNA-seq;2.ATAC-seq寻找先锋转录因子(pioneer factor, master regulator),然后进行ChIP-seq验证。当然,最近几年单细胞挺火的,ATAC-seq搭配scATAC-seq,scRNA-seq,snRNA-seq也是越来越常见的思路了。Tn5酶法是illumina针对DNA低起始量开发的微量建库方法,用于切细胞核时即形成了ATAC-seq,美国Stanford大学的William Greenleaf教授开发的方法。只需要两个步骤:1.对细胞核进行酶打断2.PCR扩增,整个实验非常简单,当ATAC-seq出现后染色质可及性分析迅速风靡各个实验室。
由于ATAC-seq的Tn5酶对细胞碎片比较敏感,所以一般要对细胞核进行提纯,可以选择密度梯度离心和流式分选的方法。宸谱生物研发了自己独特的buffer来进行ATAC-seq用于冷冻组织,极大的拓展了ATAC-seq在临床样本中的应用。欢迎各位老师联系宸谱生物进行合作。
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