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

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

DOI:
10.3791/60919-v

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09:41 min

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May 14, 2020

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

Chapters

  • 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

Automatic Translation

Automatic Translation

Translating ribosome affinity purification (TRAP) offers the possibility to dissect developmental programs with minimal processing of organs and tissues. The protocol yields high-quality RNA from cells targeted with a green fluorescent protein (GFP)-labeled ribosomal subunit. Downstream analysis tools, such as qRT-PCR or RNA-seq, reveal tissue and cell type-specific expression profiles.

Tags

Keywords: Ribosome Affinity Purification (TRAP)Arabidopsis ThalianaRoot DevelopmentCell Type-specificPolysome RNALow-input RNALateral Root FormationCell-cell CommunicationCell WallMechanical SignalingSeed SterilizationExogenous TreatmentNAA

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