Whole-genome chromatin profiling from limited numbers of cells using nano-ChIP-seq.
Whole-genome chromatin profiling from limited numbers of cells using nano-ChIP-seq.
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DOI：
10.1038/nprot.2011.402
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发表时间：
2011-09-29
期刊：
影响因子：
14.8
通讯作者：
中科院分区：
文献类型：
--
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染色质免疫沉淀（ChIP）结合高通量测序（ChIP-seq）已成为蛋白质-DNA相互作用的全基因组作图的金标准。然而，传统的ChIP方案需要使用大量的细胞，并且与当前高通量测序平台相关的文库制备步骤需要大量的DNA;这两个因素都排除了ChIP-seq技术应用于许多生物学上重要但罕见的细胞类型。在这里，我们描述了一种纳米ChIP-seq协议，结合了高灵敏度的小规模ChIP测定和定制的程序，用于从稀缺量的ChIP DNA生成高通量测序文库。在所需细胞数量方面，该方法比传统的ChIP-seq方法提高了两到三个数量级，整个过程可以在4天内完成。
Chromatin immunoprecipitation (ChIP) combined with high-throughput sequencing (ChIP-seq) has become the gold standard for whole-genome mapping of protein-DNA interactions. However, conventional ChIP protocols necessitate the use of large numbers of cells, and library preparation steps associated with current high-throughput sequencing platforms require substantial amounts of DNA; both of these factors preclude the application of ChIP-seq technology to many biologically important but rare cell types. Here we describe a nano-ChIP-seq protocol that combines a high-sensitivity small-scale ChIP assay and a tailored procedure for generating high-throughput sequencing libraries from scarce amounts of ChIP DNA. In terms of the numbers of cells required, the method provides two to three orders of magnitude of improvement over the conventional ChIP-seq method and the entire procedure can be completed within 4 d.
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影响因子：
14.8
作者：
Dahl, John Arne;Collas, Philippe
通讯作者：
Collas, Philippe
影响因子：
48
作者：
Goren, Alon;Ozsolak, Fatih;Shoresh, Noam;Ku, Manching;Adli, Mazhar;Hart, Chris;Gymrek, Melissa;Zuk, Or;Regev, Aviv;Milos, Patrice M.;Bernstein, Bradley E.
通讯作者：
Bernstein, Bradley E.
影响因子：
5.2
作者：
Dahl, John Arne;Collas, Philippe
通讯作者：
Collas, Philippe
影响因子：
30.8
作者：
Lieb, JD;Liu, XL;Brown, PO
通讯作者：
Brown, PO
影响因子：
2.7
作者：
Acevedo, Luis G.;Iniguez, A. Leonardo;Farnham, Peggy J.
通讯作者：
Farnham, Peggy J.