JoVE Journal

Developmental Biology

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

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.

Chapters in this video

0:04

Introduction

1:08

Propagation and Sterilization

1:36

Plating

2:20

Tissue Treatment (Optional)

2:51

Harvesting

3:39

Tissue Grinding and Homogenization

4:39

Total RNA Sample Collection, Crude Extract Clearing, Bead Washing

5:34

Immunoprecipitation

6:49

Lab Supply Reconditioning

7:38

Results: Representative Cell Type-Specific GFP Expression and Translational Ribosome Affinity Purification (TRAP)/Polysome RNA Quality Assessment

8:47

Conclusion

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