Why is there background in open chromatin in my sequencing data?

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Background in open chromatin may indicate digestion of accessible chromatin by pAG-MNase, due to prolonged incubation and/or incorrect temperature. However, it can also indicate over-sequencing of CUT&RUN libraries.

To resolve this:

  • Sequence libraries to 3-8 million reads.

  • Repeat assay using fresh buffers.

  • Ensure pAG-MNase digestion is performed at 4˚C.