|
Author:
Date:
2016-06-20
[Abstract] Chromatin immunoprecipitation followed by deep sequencing (ChIP-Seq) is an important strategy to study gene regulation. When availability of cells is limited, however, it can be useful to focus on specific genes to investigate in depth the role of transcription factors or histone marks. Unfortunately, performing ChIP experiments to study transcription factors’ binding to DNA can be difficult when biological material is restricted. This protocol describes a robust method to perform μChIP for over-expressed or endogenous transcription factors using ~100,000 cells per ChIP experiment (Masserdotti et al., 2015). We also describe optimization steps, which we think are critical for this protocol to work and which can be used to further reduce the number of cells.
[摘要] 染色质免疫沉淀后深层测序（ChIP-Seq）是研究基因调控的重要策略。 然而，当细胞的可用性有限时，可能有用的是专注于特定的基因深入研究转录因子或组蛋白标记的作用。 不幸的是，当生物材料受到限制时，进行ChIP实验以研究转录因子与DNA的结合可能是困难的。 该方案描述了使用约100,000个细胞/ChIP实验对过表达或内源性转录因子进行μChIP的稳健方法（Masserdotti等人，2015）。 我们还描述了优化步骤，我们认为这是协议工作的关键，可以用于进一步减少单元格的数量。
|