Geisberg Joseph V, Struhl Kevin
Harvard Medical School, Boston, Massachusetts, USA.
Curr Protoc Mol Biol. 2005 May;Chapter 21:Unit 21.8. doi: 10.1002/0471142727.mb2108s70.
Sequential Chromatin Immunoprecipitation (SeqChIP) is a powerful technique for analyzing the simultaneous association of two different proteins with genomic DNA sequences in vivo. Cellular Protein-DNA complexes are cross-linked with formaldehyde (UNIT ), and are purified via two successive immunoprecipitations, with each immunoprecipitation targeting a different protein. Protein-DNA cross-links are then reversed and DNA sequences of interest are analyzed by quantitative PCR. At each genomic region, calculated SeqChIP co-occupancy values are compared to occupancy values of singly immunoprecipitated samples. The extent of enrichment brought about by the second immunoprecipitation relative to the singly immunoprecipitated sample is directly correlated with the degree of co-occupancy between the two proteins at the genomic location assayed. In principle, the technique is not limited to Saccharomyces cerevisiae. Cells from a wide variety of organisms can be used.
序列染色质免疫沉淀法（SeqChIP）是一种强大的技术，用于分析体内两种不同蛋白质与基因组DNA序列的同时结合。细胞蛋白质-DNA复合物用甲醛交联（单元），并通过两次连续的免疫沉淀进行纯化，每次免疫沉淀针对一种不同的蛋白质。然后逆转蛋白质-DNA交联，并通过定量PCR分析感兴趣的DNA序列。在每个基因组区域，将计算出的SeqChIP共占据值与单免疫沉淀样品的占据值进行比较。第二次免疫沉淀相对于单免疫沉淀样品所带来的富集程度与所检测基因组位置上两种蛋白质的共占据程度直接相关。原则上，该技术不限于酿酒酵母。可以使用来自多种生物体的细胞。
