Description

This track represents binding events for the GA-binding protein as assayed by chromatin immunoprecipation on Affymetrix whole genome tiling 2.0R arrays in various cell types.

Methods

Chromatin IP was performed as described in Johnson et al. (2007) using the monoclonal GABPa antibody (G1, Santa Cruz Biotechnology). Libraries were prepared from the ChIP DNA using the standard Affymetrix amplification protocol. The amplified ChIP libraries were then fractionated and hybridized to human Affymetrix whole genome 2.0R arrays in biological triplicate.

The data represent peaks based on the array measurements and called by the MAT software (Johnson et al., 2006). The following settings were used: BandWidth = 300, MaxGap = 300, MinProbe = 10, Trim = 0.1, pvalue=1e-5.

Verification

These data were performed in biological triplicate. The authors performed Western blots to test the specificity of the antibody, gel electrophoresis to test the proper shearing of chromatin, and qPCR assays of several loci before hybridization to arrays.

Credits

Myers Group: David Johnson, Betsy Anton, Loan Nguyen, Cat Medina, Richard Myers.

References

Johnson DS, Mortazavi A, Myers RM, Wold B. Genome-Wide Mapping of in Vivo Protein-DNA Interactions. Science 2007 June;316:1497-1502.

Johnson WE, Li W, Meyer CA, Gottardo R, Carroll JS, Brown M, Liu XS. Model-based analysis of tiling-arrays for ChIP-chip. Proc. Natl. Acad. Sci.2006;103:12457-62.