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Microarray-based amplification and detection of RNA by nucleic acid sequence based amplification.
Mader, Andreas; Riehle, Ulrike; Brandstetter, Thomas; Stickeler, Elmar; zur Hausen, Axel; Rühe, Jürgen.
Afiliação
  • Mader A; Institute of Pathology, Freiburg University Medical Center, Breisacher Strasse 115a, 79106 Freiburg, Germany.
Anal Bioanal Chem ; 397(8): 3533-41, 2010 Aug.
Article em En | MEDLINE | ID: mdl-20596698
Nucleic acid sequence based amplification (NASBA) is a versatile in vitro nucleic acid amplification method. In this work, RNA amplification and labeling by NASBA and microarray analysis are combined in a one-step process. The NASBA reaction is performed in direct contact with capture probes. These probes are bound to surface-attached hydrogel spots generated at the chip surfaces by using a simple printing and UV irradiation process. Five gene expression and SNP parameters with known relevance in breast cancer diagnostics were chosen to demonstrate that multiplex NASBA-on-microarray analysis is possible. A minimum amount of 10 pg of total RNA was shown to be sufficient for the detection of the reference parameter RPS18, which demonstrates that the detection limit of the microarray-based NASBA assays theoretically allows single-cell assays to be performed.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA / Análise de Sequência com Séries de Oligonucleotídeos / Replicação de Sequência Autossustentável Tipo de estudo: Diagnostic_studies / Evaluation_studies Limite: Female / Humans Idioma: En Revista: Anal Bioanal Chem Ano de publicação: 2010 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: RNA / Análise de Sequência com Séries de Oligonucleotídeos / Replicação de Sequência Autossustentável Tipo de estudo: Diagnostic_studies / Evaluation_studies Limite: Female / Humans Idioma: En Revista: Anal Bioanal Chem Ano de publicação: 2010 Tipo de documento: Article