RESUMO
Chromatin-associated RNA (caRNA) is a newly identified class of RNA molecules stably associated with chromatin, maintaining the higher order structure of euchromatic regions in an accessible form (Schubert et al., Mol Cell, doi:10.1016/j.molcel.2012.08.021, 2012). This section provides a detailed protocol describing the isolation of small caRNA from Drosophila cells and preparation of libraries suited for stranded small (20-200 bp) RNA deep sequencing on the Illumina platform.
Assuntos
Cromatina/metabolismo , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala/métodos , RNA Nuclear Pequeno/genética , RNA Nuclear Pequeno/isolamento & purificação , Análise de Sequência de RNA/métodos , Animais , Drosophila melanogaster/metabolismoRESUMO
Chromatin-associated RNA (caRNA) is a newly identified class of RNA stably linked to chromatin and responsible for maintaining the higher order structure of euchromatic regions in an accessible form (Schubert et al., Mol Cell 48, 434 - 444, 2012). This section provides a pipeline of bioinformatic analysis for this specific type of RNA-Seq. It can be run locally by combining several scripts or be carried out on the Galaxy platform (Giardine et al., Genome Res 15:1451-1455, 2005).