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dc.contributor.authorScheler, Ott
dc.contributor.authorGlynn, Barry
dc.contributor.authorParkel, Sven
dc.contributor.authorPalta, Priit
dc.contributor.authorToome, Kadri
dc.contributor.authorKaplinski, Lauris
dc.contributor.authorRemm, Maido
dc.contributor.authorMaher, Majella
dc.contributor.authorKurg, Ants
dc.date.accessioned2018-09-20T16:24:01Z
dc.date.available2018-09-20T16:24:01Z
dc.date.issued2009-01-01
dc.identifier.citationScheler, Ott; Glynn, Barry; Parkel, Sven; Palta, Priit; Toome, Kadri; Kaplinski, Lauris; Remm, Maido; Maher, Majella; Kurg, Ants (2009). Fluorescent labeling of nasba amplified tmrna molecules for microarray applications. BMC Biotechnology 9 ,
dc.identifier.issn1472-6750
dc.identifier.urihttp://hdl.handle.net/10379/13814
dc.description.abstractBackground: Here we present a novel promising microbial diagnostic method that combines the sensitivity of Nucleic Acid Sequence Based Amplification (NASBA) with the high information content of microarray technology for the detection of bacterial tmRNA molecules. The NASBA protocol was modified to include aminoallyl-UTP (aaUTP) molecules that were incorporated into nascent RNA during the NASBA reaction. Post-amplification labeling with fluorescent dye was carried out subsequently and tmRNA hybridization signal intensities were measured using microarray technology. Significant optimization of the labeled NASBA protocol was required to maintain the required sensitivity of the reactions. Results: Two different aaUTP salts were evaluated and optimum final concentrations were identified for both. The final 2 mM concentration of aaUTP Li-salt in NASBA reaction resulted in highest microarray signals overall, being twice as high as the strongest signals with 1 mM aaUTP Na-salt. Conclusion: We have successfully demonstrated efficient combination of NASBA amplification technology with microarray based hybridization detection. The method is applicative for many different areas of microbial diagnostics including environmental monitoring, bio threat detection, industrial process monitoring and clinical microbiology.
dc.publisherSpringer Nature
dc.relation.ispartofBMC Biotechnology
dc.subjectsequence-based amplification
dc.subjectDNA microarrays
dc.subjectrna
dc.subjectexpression
dc.subjectprobes
dc.subjecttarget
dc.subjectassay
dc.titleFluorescent labeling of nasba amplified tmrna molecules for microarray applications
dc.typeArticle
dc.identifier.doi10.1186/1472-6750-9-45
dc.local.publishedsourcehttps://bmcbiotechnol.biomedcentral.com/track/pdf/10.1186/1472-6750-9-45?site=bmcbiotechnol.biomedcentral.com
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