| cDNA Library Construction for Next-Generation Sequencing to Determine the Transcriptional Landscape of Legionella pneumophila. | |
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MedLine Citation:
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PMID: 23150420 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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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. |
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Authors:
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Tobias Sahr; Carmen Buchrieser |
Publication Detail:
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Type: JOURNAL ARTICLE |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 954 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2013 |
Date Detail:
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Created Date: 2012-11-14 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: - |
Other Details:
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Languages: ENG Pagination: 555-566 Citation Subset: - |
Affiliation:
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Biologie des Bactéries Intracellulaires, Institut Pasteur, Paris, France. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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