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ChIP-qPCR Negative Control Primers

SABiosciences' ChIP-qPCR Negative Control Primers measure the relative amount of non-specific genomic DNA that co-precipitates during the ChIP procedure. The limited amount of highly enriched ChIP DNA material and the very sensitive nature of real-time PCR detection demand comprehensive negative controls to insure specificity of target IP signals. The binding of a significant amount of non-specific genomic DNA to the target-specific antibody falsely elevates qPCR signal intensities and the apparent amount of nuclear factor binding. The ideal solution is to detect DNA that is not associated with a target factor binding site. However, biological variables make the choice of that sequence challenging, because many genes and promoters overlap with other genes or promoters and their nuclear factor binding sties.

The ChIP-qPCR Negative Control Primers detect specific genomic DNA sequences within ORF-free intergenic regions or "promoter deserts" lacking any known or predicted structural genes. The isolation of these regions minimizes the likelihood of promoter-specific nuclear factor interactions. The threshold cycle (Ct) values from specific ChIP fractions and these primers should be identical to the negative ChIP control (non-immune serum fraction). This result indicates similar levels of non-specific genomic DNA co-precipitation between ChIP fractions. Because each antibody and its corresponding non-immune serum can interact differently and to varying extents with non-specific DNA, a negative control should be performed for every experimental ChIP fraction. Include the ChIP-qPCR Negative Control Primers in your ChIP experiment to more accurately assess the specificity of your target binding enrichment.

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