ChIP-Seq Analysis in Neurospora crassa.

ChIP-Seq Analysis in Neurospora crassa.
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粗糙脉孢菌的 ChIP-Seq 分析。

DOI:
10.1007/978-1-4939-7804-5_19
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发表时间:
2018
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Lewis,ZacharyA
Lewis,ZacharyA
中科院分区:
--
文献类型:
--
作者:
Ferraro,AileenR;Lewis,ZacharyA

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染色质免疫沉淀与下一代测序(ChIP-seq)配对可用于确定转录因子、转录机制或组蛋白修饰的全基因组分布。DNA-蛋白质相互作用通过加入甲醛共价交联。染色质被制备和剪切,然后用适当的抗体免疫沉淀。在交联逆转和用蛋白酶处理后,对所得DNA片段进行测序并映射到参考基因组以确定总体富集。在这里,我们描述了一种ChIP-seq的方法来研究丝状真菌粗糙脉孢菌中的蛋白质-DNA相互作用。
Chromatin immunoprecipitation paired with next-generation sequencing (ChIP-seq) can be used to determine genome-wide distribution of transcriptions factors, transcriptional machinery, or histone modifications. DNA–protein interactions are covalently cross-linked with the addition of formaldehyde. Chromatin is prepared and sheared, then immunoprecipitated with the appropriate antibody. After reversal of cross-linking and treating with protease, the resulting DNA fragments are sequenced and mapped to the reference genome to determine overall enrichment. Here we describe a method of ChIP-seq for investigating protein–DNA interactions in the filamentous fungusNeurospora crassa.
DOI: 10.1038/nmeth.1923
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