Real-time stability testing of air-dried primers and fluorogenic hydrolysis probes stabilized by trehalose and xanthan

Real-time stability testing of air-dried primers and fluorogenic hydrolysis probes stabilized by trehalose and xanthan

A method for conserving primers and differently labeled fluorogenic hydrolysis (i.e., TaqMan) probes at ambient conditions is presented. Primers and hydrolysis probes with four different fluorophore-quencher combinations (6- FAM-BHQ1, HEX-BHQ1, ROX-BHQ650, and Cy5-BHQ2) were mixed with trehalose and...

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Veröffentlicht in:BioTechniques. - 1988. - 57(2014), 3 vom: 11., Seite 151-5
1. Verfasser: Rombach, Markus (VerfasserIn)
Weitere Verfasser: Kosse, Dominique, Faltin, Bernd, Wadle, Simon, Roth, Günter, Zengerle, Roland, von Stetten, Felix
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2014
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Journal Article Research Support, Non-U.S. Gov't air-drying preservation primer probe stability testing trehalose xanthan DNA Primers mehr... Excipients Fluorescent Dyes Polysaccharides, Bacterial Trehalose B8WCK70T7I xanthan gum TTV12P4NEE
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500 |a ErratumIn: Biotechniques. 2014 Oct;57(4):213 
500 |a Citation Status MEDLINE 
520 |a A method for conserving primers and differently labeled fluorogenic hydrolysis (i.e., TaqMan) probes at ambient conditions is presented. Primers and hydrolysis probes with four different fluorophore-quencher combinations (6- FAM-BHQ1, HEX-BHQ1, ROX-BHQ650, and Cy5-BHQ2) were mixed with trehalose and xanthan at final concentrations of 56 mM and 2.78 mM, respectively. Mixtures were air-dried at 23°C for 30 min on strips composed of cyclo olefin polymer (COP), a material widely used in the manufacturing of in vitro diagnostic (IVD) test carriers. After one year of storage, the functionality of the primers and fluorophore-quencher combinations was validated by real-time polymerase chain reaction (real-time PCR), confirming their stability when stored in the presence of stabilizers, with the best results achieved using trehalose. This approach could be of great benefit for manufacturing IVD systems, for example, for genotyping applications based on multiplexing using different fluorescent dyes 
650 4 |a Journal Article 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a air-drying 
650 4 |a preservation 
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650 4 |a stability testing 
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650 7 |a Fluorescent Dyes  |2 NLM 
650 7 |a Polysaccharides, Bacterial  |2 NLM 
650 7 |a Trehalose  |2 NLM 
650 7 |a B8WCK70T7I  |2 NLM 
650 7 |a xanthan gum  |2 NLM 
650 7 |a TTV12P4NEE  |2 NLM 
700 1 |a Kosse, Dominique  |e verfasserin  |4 aut 
700 1 |a Faltin, Bernd  |e verfasserin  |4 aut 
700 1 |a Wadle, Simon  |e verfasserin  |4 aut 
700 1 |a Roth, Günter  |e verfasserin  |4 aut 
700 1 |a Zengerle, Roland  |e verfasserin  |4 aut 
700 1 |a von Stetten, Felix  |e verfasserin  |4 aut 
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773 1 8 |g volume:57  |g year:2014  |g number:3  |g day:11  |g pages:151-5 
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