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Ligation-free ribosome profiling of cell type-specific translation in the brain

Ribosome profiling has emerged as a powerful tool for genome-wide measurements of translation, but library construction requires multiple ligation steps and remains cumbersome relative to more conventional deep-sequencing experiments. We report a new, ligation-free approach to ribosome profiling tha...

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Detalles Bibliográficos
Autores principales: Hornstein, Nicholas, Torres, Daniela, Das Sharma, Sohani, Tang, Guomei, Canoll, Peter, Sims, Peter A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4934013/
https://www.ncbi.nlm.nih.gov/pubmed/27380875
http://dx.doi.org/10.1186/s13059-016-1005-1
Descripción
Sumario:Ribosome profiling has emerged as a powerful tool for genome-wide measurements of translation, but library construction requires multiple ligation steps and remains cumbersome relative to more conventional deep-sequencing experiments. We report a new, ligation-free approach to ribosome profiling that does not require ligation. Library construction for ligation-free ribosome profiling can be completed in one day with as little as 1 ng of purified RNA footprints. We apply ligation-free ribosome profiling to mouse brain tissue to identify new patterns of cell type-specific translation and test its ability to identify translational targets of mTOR signaling in the brain. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13059-016-1005-1) contains supplementary material, which is available to authorized users.