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==Important Notes== *The addition of 4X the LIS amount during depletion results in an extremely viscous solution. It is unclear whether the LIS lyses nuclei. This viscous solution prevents the centrifugation of nuclei and the LIS reagent couldn't be removed *The depleted nuclei may need to be centrifuged at a much higher speed than undepleted nuclei based on https://www.ncbi.nlm.nih.gov/pmc/articles/PMC148640/pdf/273779.pdf Depleted nuclei were centrifuged at 6000g for 15-20 minutes *The paper also suggests that the LIS "extracts" the histone binding proteins from the nuclei causing a DNA halo of DNA strands that permeate out of the nuclei, but do not actually rupture the nuclei. The picture in the publication needs to be inserted here.
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