Nov 06, 2025

Public workspaceMouse Harvest for Single Nuclear Sequencing of Liver

DOI
https://dx.doi.org/10.17504/protocols.io.bp2l6zerdgqe/v1
  • Daniel Alligood1,
  • Alex Park1,
  • Jun Guo1
  • 1Department of Surgery Washington University in St. Louis
Daniel Alligood
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DOI: https://dx.doi.org/10.17504/protocols.io.bp2l6zerdgqe/v1
Protocol Citation: Daniel Alligood, Alex Park, Jun Guo 2025. Mouse Harvest for Single Nuclear Sequencing of Liver. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6zerdgqe/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 01, 2025
Last Modified: November 07, 2025
Protocol Integer ID: 231272
Keywords: tissue collection, single nuclear sequencing of liver, single nuclear sequencing, standard murine tissue collection method, standard murine tissue collection method in preparation, liver this protocol, liver
Abstract
This protocol is our standard murine tissue collection method in preparation for single nuclear sequencing of liver.
Troubleshooting
Tube Preparation
Annotate RNAase free 1.5ml Eppendorf tubes with mouse ID. Create "A" and "B" duplicates

Fill tubes with 500uL of RNALater (Invitrogen AM7021)
Mouse Preparation
Fast mice for 4 hours before harvest
Induce anesthesia with isoflurane
Tissue Collection
Create a midline laparotomy
Divide the falciform ligament and remove the liver en-bloc
Section off the left anterior lobe
Sharply section two 0.5x0.5cm liver pieces and place in the RNALater tubes. Place the remaining portion of the lobe in a cassette and fix with 10% formalin.
Flash freeze the tubes with liquid nitrogen and store in the -80C freezer until time for single nuclear isolation.