Abstract:
: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.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Rombach M,Kosse D,Faltin B,Wadle S,Roth G,Zengerle R,von Stetten Fdoi
10.2144/000114207subject
Has Abstractpub_date
2014-09-01 00:00:00pages
151-5issue
3eissn
0736-6205issn
1940-9818pii
000114207journal_volume
57pub_type
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