Sufficient reagents are provided to perform three sets of chromatin sonication optimization experiments. Each set would include five different conditions for sonication to optimize the parameters. Reagents are provided for a total of 15 reactions, including preparation of cell lysates from 107 cells/reaction, nuclei lysis, sonication, cross-link reversal and DNA purification.
General description
The Imprint Ultra Chromatin Optimization Kit provides reagents needed to optimize chromatin sonication/shearing conditions before performing ChIP. It is imperative that chromatin is completely sheared into fragments with a peak around 500 base pairs (bp) for ChIP-qPCR assays, and ﹤ 500 bp for ChIP-Seq and ChIP-chip, for maximum specificity and resolution, and to ensure low background. For higher resolution applications like genome-wide location analysis by ChIP-chip or ChIP-Seq, sonication of input chromatin to below 500 bp, with a peak around 250 bp, is critical.
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