Rice Meiotic Chromosome Spread Preparation of Pollen Mother Cells
从水稻花粉母细胞中分离与制备处于减数分裂期细胞染色体
发布: 2014年07月20日第4卷第14期 DOI: 10.21769/BioProtoc.1189 浏览次数: 12234
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参见作者原研究论文
The authors used this protocol in:
Oct 2013
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Abstract
In this protocol, we describe a simple and efficient method for meiotic chromosome spread preparation in rice pollen mother cells. Meiotic chromosome preparation by spreading itself is an important technique for plant cytogenetics (Higgins et al., 2004; Chelysheva et al., 2012; Wang et al., 2009); furthermore, it is a crucial step for applying other cytogenetic methods including Fluorescence in situ hybridization (FISH) and immunostaining.
Materials and Reagents
- Young panicles containing meiocytes of rice (Oryza sativa ssp japonica cv. Zhonghua 11)
- 70%, 90% and 100% ethanol (Analytical Reagents)
- 60% (v/v) acetic acid (Analytical Reagents)
- Carmine (Sigma-Aldrich, catalog number: C1022-25G )
- Liquid nitrogen
- Vectashield Mounting Medium with DAPI (Vector Laboratories, catalog number: H-1200 )
- Carnoy’s fixative (see Recipes)
- Aceto-carmine (see Recipes)
Equipment
- Stereo microscope
- Fluorescence microscope (Zeiss, model: AX10 )
- Microscopic slides and cover slips
- Dissection needles and fine forceps
- Heating block
- CCD camera (Hamamatsu Photonics K.K., model: ORCA-R2 C10600 )
Procedure
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版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Li, X. and Wu, C. (2014). Rice Meiotic Chromosome Spread Preparation of Pollen Mother Cells. Bio-protocol 4(14): e1189. DOI: 10.21769/BioProtoc.1189.
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分类
植物科学 > 植物分子生物学 > DNA > DNA 结构
植物科学 > 植物细胞生物学 > 细胞成像
分子生物学 > DNA > DNA 结构
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