TRAP-Seq

Targeted Purification of Polysomal mRNA

Targeted purification of polysomal mRNA (TRAP-Seq) maps translating mRNAs under various conditions (Jiao et al., 2010). In this method, tagged ribosomal proteins are expressed in cells. The tagged ribosomal proteins are purified and the RNA isolated. The RNA is reverse-transcribed to cDNA. Deep sequencing of the cDNA provides single-base resolution of translating RNAs.

Advantages:

• Allows detection of translating RNAs
• RNAs translated by specific targeted ribosomes can be assessed
• No prior knowledge of the RNA is required
• Provides a genome-wide RNA screen

Disadvantages:

• Not as specific as more recently developed methods, such as Ribo-Seq

Reagents:

Illumina Library prep and Array Kit Selector



Reviews:

Nussbacher J. K., Batra R., Lagier-Tourenne C. and Yeo G. W. RNA-binding proteins in neurodegeneration: Seq and you shall receive. Trends Neurosci. 2015;38:226-236

Maze I., Shen L., Zhang B., et al. Analytical tools and current challenges in the modern era of neuroepigenomics. Nat Neurosci. 2014;17:1476-1490



References:

Reynoso M. A., Juntawong P., Lancia M., Blanco F. A., Bailey-Serres J. and Zanetti M. E. Translating Ribosome Affinity Purification (TRAP) followed by RNA sequencing technology (TRAP-SEQ) for quantitative assessment of plant translatomes. Methods Mol Biol. 2015;1284:185-207

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History: TRAP-Seq

Revision by sbrumpton on 2017-06-21 07:50:25 - Show/Hide