Materials and Reagents

Materials
1. 200 μL pipette tips
2. 1 mL pipette tips
3. 5 mL pipette tips
4. 100-μm Nylon cell stainer (Corning Incorporated, FALCON, catalog number: 352350)
5. 40-μm Nylon cell stainer (Corning Incorporated, FALCON, catalog number: 352340)
6. 50 mL centrifuge tube
7. 1.5 mL centrifuge tube
8. Arabidopsis seedlings
9. Liquid nitrogen
Reagents
1. Sucrose (Sinopharm, SCR, catalog number: 10021418)
2. Ficoll 400 (Sigma-Aldrich, catalog number: F9378-5G)
3. Dextran T40 (Solarbio, catalog number: D8250)
4. Tris (Sangon Biotech, catalog number: A610195)
5. HCl (Sinopharm, SCR, catalog number: 10011018)
6. MgCl₂ (RHAWN, catalog number: R007504-500 g)
7. Triton X-100 (Sigma-Aldrich, catalog number: V900502-100ML)
8. phenylmethylsulfonyl fluoride (PMSF) (Sigma-Aldrich, catalog number: PMSF-RO)
9. β-mercaptoethanol (Sigma-Aldrich, catalog number: 07604-100ML)
10. Protease inhibitor cocktail (Roche, cOmplete Tablets EDTA-free, EASYpack, catalog number: 04693132001)
11. KCl (Sinopharm, SCR, catalog number: 10016318)
12. HEPES (Sigma-Aldrich, catalog number: H3375)
13. Hexylene glycol (Sangon Biotech, catalog number: A607060)
14. Spermine (Sigma-Aldrich, catalog number: S3256)
15. Spermidine (Sigma-Aldrich, catalog number: S2626)
16. Glycerol (Sangon Biotech, catalog number: A100854)
17. NaCl (Sinopharm, SCR, catalog number: 10019318)
18. 0.5 M EDTA, pH 8.0 (Biosharp, catalog number: 69075458)
19. Dithiothreitol (DTT) (Beyotime, catalog number: ST043-5g)
20. Urea (Sangon Biotech, catalog number: A600148)
21. Nonidet P-40 (NP-40) (Roche, catalog number: 11332473001)
22. Anti-Histone H3 antibody (Abcam, catalog number: ab18521)
Stock solution
Stock solutions for the buffers can be made before and stored at 4°C (1-5) or -20°C (6-10). On the day of the experiment, the buffers can be diluted as required.
1. 1 M Tris-HCl, pH 7.4
2. 20% Triton X-100 (v/v)
3. 20% NP-40 (v/v)
4. 2 M KCl
5. 0.5 M HEPES-KOH, pH 7.0
6. 50 mM Spermine
7. 125 mM Spermidine
8. 10 mM PMSF
9. 1.0 M DTT
10. Protease inhibitor cocktail (100×) (see Recipes)
11. Honda buffer (see Recipes)
12. Nuclei isolation buffer (NIB) buffer (see Recipes)
13. Glycerol buffer (see Recipes)
14. Nuclei lysis buffer (see Recipes)

Equipment

Centrifuge (ThermoFisher, catalog number: 75004380)
Vortex shaker
Mortar

Procedure