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. 2001 Aug 15;15(16):2069–2082. doi: 10.1101/gad.906601

Figure 2.

Figure 2

Quantification of chromatin immunoprecipitation by real-time PCR. Chromatin extracted from cross-linked Rat1 or myc−/− cells was immunoprecipitated with anti-Myc antibodies. DNA was recovered and used as a template for real-time PCR in a Taqman 5700 (Perkin Elmer). (A) Representative PCR-amplification curves, as displayed by the Taqman 5700. In this example, we used primers amplifying the NUC E-box domain (amplicon +574, Fig. 1). Calculation of the amount of immunoprecipitated NUC E-box DNA relative to that present in total input chromatin is shown at the bottom. (CT) Cycle threshold, cycle number at which each PCR reaction reaches a predetermined fluorescence threshold, set within the linear range of all reactions. (B) Graphic representation of the data for NUC and control promoters analyzed in the same experiment (GLU, GLY, PCNA, and ACHR). In this experiment, cells were cross-linked and harvested 4 h after mitogenic stimulation.