Nuclear Extraction from Arabidopsis thaliana
拟南芥中细胞核的分离
发布: 2012年12月20日第2卷第24期 DOI: 10.21769/BioProtoc.306 浏览次数: 34216
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参见作者原研究论文
The authors used this protocol in:
Jun 2012
Abstract
This protocol is to isolate nuclei from Arabidopsis cells. They can be further used for other experiments, such as nuclear protein detection, nuclear protein immunoprecipitation and so on.
Materials and Reagents
- Tris-HCl (pH 7.4)
- Glycerol
- KCl
- EDTA (pH 7.5)
- MgCl2
- Sucrose
- Triton X-100
- Murashige and Skoog basal medium (Sigma-Aldrich, catalog number: M0404-10L )
- Phenylmethanesulfonylfluoride (PMSF)
- Dithiothreitol (DTT)
- Proteinase inhibitor (PI) (complete EDTA-free) (Roche Diagnostics, catalog number: 04693132001 )
- Phytagel (Sigma-Aldrich, catalog number: P8169-1KG )
- Liquid nitrogen
- Lysis buffer (LB) (see Recipes)
- Nuclei resuspension buffer with 0.2% Triton X-100 (NRBT) (see Recipes)
- Nuclei resuspension buffer (NRB) (see Recipes)
- Nuclei storage buffer (NSB) (see Recipes)
- MS (see Recipes)
Equipment
- Centrifuges (e.g. Eppendorf centrifuge 5810 R that can be refrigerated and will hold 50 ml tubes)
- Mortar and pestle
- 100 μm and 40 μm nylon mesh (BD Biosciences, Falcon®, catalog number: REF352360 , REF352340 )
- 50 ml conical tube
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Xu, F. and Copeland, C. (2012). Nuclear Extraction from Arabidopsis thaliana. Bio-protocol 2(24): e306. DOI: 10.21769/BioProtoc.306.
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分类
植物科学 > 植物细胞生物学 > 细胞器分离
系统生物学 > 蛋白质组学 > 细胞核
细胞生物学 > 细胞器分离 > 细胞核
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