体外全基因组转录因子结合位点鉴定。

In Vitro Genome-Wide Identification of Transcription Factor Binding Sites.

机构信息

Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan.

出版信息

Methods Mol Biol. 2025;2869:169-182. doi: 10.1007/978-1-0716-4204-7_17.

Abstract

DNA affinity purification followed by sequencing (DAP-seq) profiles transcription factor (TF) binding sites in vitro, in which recombinant TF captures the genomic fragments with TF binding sites. Here, I describe the preparation of pre-PCR DNA-seq libraries for 96 TFs, including the expression and purification of recombinant TF, the affinity purification of TF binding fragments, and the sequencing-ready library preparation. Using this method, up to 96 DAP-seq libraries can be prepared in 2 days.

摘要

DNA 亲和纯化结合测序（DAP-seq）技术可在体外分析转录因子（TF）结合位点，在此过程中，重组 TF 可捕获含有 TF 结合位点的基因组片段。本文描述了 96 种 TF 的预 PCR DNA-seq 文库的制备方法，包括重组 TF 的表达和纯化、TF 结合片段的亲和纯化以及测序文库的准备。使用该方法，在 2 天内可制备多达 96 个 DAP-seq 文库。