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URL: https://dx.doi.org/DOI:10.17504/protocols.io.bs99nh96

Authors: Luciano Martelotto

Group: Human Cell Atlas Method Development Community, Single Cell Core, Harvard Medical School

Summary: This protocol is the result of the combination of various nuclei isolation protocols for single cell RNA-seq experiments using droplet-based methods, and is an extension of the Frankenstein (D.O.F means Daughter Of Frankenstein). Developed to prepare nuclei isolates from fresh and frozen material of small-to-large sizes. The good thing is that it does not uses FACS but OptiPrep® continuous gradient to remove debris. It is the alternative protocol when FACS is not available. This protocol works better with smaller samples. After centrifugation the nuclei will form a pellet and contaminating membranes will float at the top. This continuous gradient can be applied for as many times as needed to the same prep to continue cleaning up your nuclei suspensions.

Affiliations: Harvard Medical School

External URL: https://singlecellcore.hms.harvard.edu/

Version: 2

Publication Date: 2021

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