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. Author manuscript; available in PMC: 2018 Mar 26.
Published in final edited form as: Bio Protoc. 2017 Dec 20;7(24):e2654. doi: 10.21769/BioProtoc.2654

Figure 2. Pre-ultracentrifugation preparation.

Figure 2

1. Using scissors mince collected muscles in the tube. Intact muscles are shown on the left and minced muscles on the right. 2. Suspend the minced muscles in 10 ml homogenization buffer. 3. Homogenize muscle using a Dounce homogenizer. 4. Pass homogenate through a 40 μm filter into a 50 ml conical tube. Incompletely disrupted tissue and large debris will remain in the filter. 5. Centrifuge the filtrate to obtain a crude nuclear pellet (arrowhead). DAPI clearly labels the nuclear pellet under UV light. Warning: when homogenizing a vacuum may form between the pestle and homogenate. If the pestle is sucked down into the homogenizer, it can strike with sufficient force to shatter the homogenizer.