cDNA library construction for next-generation sequencing to determine the transcriptional landscape of Legionella pneumophila.
Methods Mol Biol
; 954: 555-66, 2013.
Article
em En
| MEDLINE
| ID: mdl-23150420
ABSTRACT
The adaptation of Legionella pneumophila to the different conditions it encounters in the environment and in the host is governed by a complex regulatory system. Current knowledge of these regulatory networks and the transcriptome responses of L. pneumophila is mainly based on microarray analysis and limited to transcriptional products of annotated protein-coding genes. The application of the Next-Generation Sequencing (NGS) technology allows now genome-wide strand-specific sequencing and accurate determination of all expressed regions of the genome to reveal the complete transcriptional network and the dynamic interplay of specific regulators on a genome-wide level. NGS-based techniques promote deeper understanding of the global transcriptional organization of L. pneumophila by identifying transcription start sites (TSS), alternative TSS and operon organization, noncoding RNAs, antisense RNAs, and 5'-/3'-untranslated regions. In this chapter we describe the construction of cDNA libraries for (1) RNA deep sequencing (RNA-seq) and (2) TSS mapping using the Illumina technology.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Biblioteca Gênica
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Legionella pneumophila
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Sequenciamento de Nucleotídeos em Larga Escala
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Transcriptoma
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article