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Biologia do Desenvolvimento

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翻译核糖体亲和纯化（TRAP）以研究细胞类型特异性尺度下阿拉伯生物基塔利亚纳根发育

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Biologia do Desenvolvimento

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JoVE Journal Biologia do Desenvolvimento

Translating Ribosome Affinity Purification (TRAP) to Investigate Arabidopsis thaliana Root Development at a Cell Type-Specific Scale

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翻译核糖体亲和纯化（TRAP）以研究细胞类型特异性尺度下阿拉伯生物基塔利亚纳根发育

DOI:

10.3791/60919-v

•

09:41 min

•

May 14, 2020

•

Martha Thellmann, Tonni Grube Andersen, Joop EM Vermeer³

¹Department of Plant and Microbial Biology,University of Zurich, ²Biophore – UNIL, ³Laboratory of Cell and Molecular Biology, Institute of Biology,University of Neuchâtel

Capítulos

00:04Introduction
01:08Propagation and Sterilization
01:36Plating
02:20Tissue Treatment (Optional)
02:51Harvesting
03:39Tissue Grinding and Homogenization
04:39Total RNA Sample Collection, Crude Extract Clearing, Bead Washing
05:34Immunoprecipitation
06:49Lab Supply Reconditioning
07:38Results: Representative Cell Type-Specific GFP Expression and Translational Ribosome Affinity Purification (TRAP)/Polysome RNA Quality Assessment
08:47Conclusion

Summary

Tadução automática

翻译核糖体亲和纯化（TRAP）提供了以最少的器官和组织处理来解剖发育方案的可能性。该协议从绿色荧光蛋白（GFP）标记的核糖体亚组细胞中产生高质量的RNA。下游分析工具（如 qRT-PCR 或 RNA-seq）可显示组织和细胞类型特定的表达特征。

Tags

Ribosome Affinity Purification (TRAP)Arabidopsis Thaliana Root Development Cell Type-specific Polysome RNA Low-input RNA Lateral Root Formation Cell-cell Communication Cell Wall Mechanical Signaling Seed Sterilization Exogenous Treatment NAA

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