Myelin removal step for Isolation of Nuclei from Adult Human Brain Tissue for 10x Genomics Platform

Science
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The Allen Institute has published laboratory protocols on protocol.io to enable the scientific community to utilize the techniques that drive our science. We received a question about myelin removal as part of our “Isolation of Nuclei from Adult Human Brain Tissue for 10x Genomics Platform” standard operating procedure: “…We have problems with the myelin removal step. We have used the removal beads from MACS and a magnetic stand from cell signaling. However, the beads won’t bind. Did you also experience this? And did you add some dynabeads too?”

We suspect others may have this same question so we plan to address it here, on our community forum, in the near future. Please stay tuned.

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We have only used the DynaMag series of magnets from Thermo Fisher. Specifically, we use the DynaMag-15 for routine preps. We do not add any Dynabeads to the preps. Increasing the concentration of the magnetic beads may help. A 1:5 dilution (beads:buffer) is useful for heavily myelinated samples. Increasing the volume of buffer used during the magnetic separation step to up to 7 ml can also help with binding. It is also worth noting that if your sample is not heavily myelinated, you can skip this myelin removal step altogether.