About ChIP on chip

Chromatin Immunoprecipitation (ChIP) is a well-established and reliable method used to identify the DNA binding site of a protein of interest such as transcription factors, histones and polymerases¹.

The combination of OGT microarrays with chromatin immunoprecipitation (ChIP) is a powerful technique that can be used to analyse and identify the positions on the genome where DNA: protein interactions occur. This has proved a powerful technique for the identification of the binding positions of global DNA binding proteins and the changes in a particular protein's binding under various environmental conditions.

In ChIP on chip the first step is the crosslinking of proteins to the genomic DNA. Following crosslinking and cell lysis the DNA is fragmented by sonication. The DNA:protein complexes are then immunoprecipitated using an antibody to the protein of interest. The protein is removed and the DNA, which will correspond to the protein of interest's binding site, can then be identified. OGT 60 mer oligonucleotide microarrays are ideal for the detection of the immunoprecipitated DNA fragments.

The DNA is labelled using incorporation of Cy-dCTP with Klenow. Following purification the labelled DNA cna then be hybidised to an OGT microarray.

¹reviewed Buck MJ and Lieb J. (2004) Genomics 83 p349-360

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