Micrococcal Nuclease (MNase) Assay of Arabidopsis thaliana Nuclei
微球菌核酸酶（Mnase）试验分析拟南芥染色质结构
发布: 2013年04月05日第3卷第7期 DOI: 10.21769/BioProtoc.455 浏览次数: 24887
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Aug 2012
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Abstract
Micrococcal nuclease (MNase) is able to produce double-strand breaks within nucleosome linker regions. The efficiency of MNase digestion depends on the degree of chromatin compaction, being more easily digested the regions of less compacted chromatin. The MNase protocol described here can be used to asses changes in the chromatin structure of nuclei extracted from Arabidopsis seedlings.
Keywords: Arabidopsis thaliana (拟南芥)
Nuclei extraction (细胞核提取)
Microccocal nuclease digestion (microccocal核酸酶消化)
Chromatin density (染色质密度)
Materials and Reagents
- Micrococcal Nuclease (MNase) (Roche Applied Science, catalog number: 10107921001 , 15,000 U)
- Proteinase K (Roche Applied Science, catalog number: 03115836001 )
- PIPES (Sigma-Aldrich, catalog number: P-9291 )
- Liquid nitrogen
- Sucrose
- KCl
- MgCl2
- CaCl2
- Triton X-100
- 1 mM PMSF (freshly added)
- Tris/HCl (pH 7.8)
- EDTA
- SDS
- Ethidium bromide
- Nuclei extraction buffer A, B, C (see Recipes)
- MNase buffer (see Recipes)
- 2x stop buffer (see Recipes)
- 10x Proteinase K buffer (see Recipes)
Equipment
- Pestle and mortar
- Centrifuges
- 37 °C oven
- 70 μm Nylon mesh
- 50 ml Falcon tubes
- Centrifuge tubes
- 2 ml microtubes
- Nylon mesh
Procedure
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文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Armengot, L. and Moreno-Romero, J. (2013). Micrococcal Nuclease (MNase) Assay of Arabidopsis thaliana Nuclei. Bio-protocol 3(7): e455. DOI: 10.21769/BioProtoc.455.
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分类
细胞生物学 > 细胞器分离 > 细胞核
分子生物学 > DNA > DNA 结构
植物科学 > 植物分子生物学 > DNA > DNA 结构
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