Question: How can I reduce myelin debris from brain in the Chromium Nuclei Isolation kit?
Answer: The presence of small debris in brain nuclei suspensions prepared with the the Nuclei Isolation kit should be expected. Debris typically observed in these samples is likely the presence of myelin, a protein that insulates neuronal axons and allows for the transmission of electrical impulses across neurons. Myelination increases during development; adult brains (mice older than 8 weeks) will have higher levels of myelin content compared to embryonic or juvenile (3-8 week) mouse brains. (https://www.frontiersin.org/articles/10.3389/fncel.2018.00424/full)
Performing an additional filtration step (30µm strainer) on adult mouse brain samples at the end of the protocol can reduce large, problematic debris without incurring a significant loss of nuclei. Small debris in the Nuclei Isolation kit is normal and expected.
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