Recipe

Nuclei Purification Buffer

In advance, prepare a 7× stock solution of cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail (Sigma-Aldrich 11836170001) and a 100 mm stock solution of spermine (e.g., Sigma-Aldrich S1141-1G) in DNase-, RNase-free H2O. Store both at −20°C until use. Prepare the recipe as follows:

1.25× Stock Solution II 3.2 mL
1 m DTT (e.g., Sigma-Aldrich 646563, once opened, store at −20°C) 8 µL
7× cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail 572 µL
100 mm Spermine 4 µL
Protector RNase inhibitor (Sigma-Aldrich 3335399001) (40 U/µL) 20 µL
DNase-, RNase-free H2O 196 µL

  • Prepare this solution in a 15-mL conical centrifuge tube (RNase/DNase-free). Because of the high viscosity of this solution, it is important to vortex it vigorously to mix thoroughly, followed by centrifugation at ∼3000 rpm for ∼30 sec at 4°C to remove air bubbles. Prepare fresh and store on ice.

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  1. doi:10.1101/pdb.rec107968 Cold Spring Harb Protoc 2022: pdb.rec107968- © 2022 Cold Spring Harbor Laboratory Press
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