Preservation of primer and probes on "ready-to-use" 96-well microtiter plates: A step forward towards enhancing throughput and harmonization of real-time PCR applications in food and feed control

Zagon,J.; Kurth,S.; Ehlers,A.; Linke,B.; Lampen,A.; Broll,H.

Three different agents, trehalose (TRH), gelatine (GEL) and polyethylene glycol (PEG) were investigated regarding their capability to preserve oligonucleotides, such as primer and probes or double-stranded plasmid DNA, pre-coated in 96-wells microtiter plates. The evaluation of the retained efficiency of selected multi-copy and two single-copy PCR (polymerase chain reaction) systems revealed best-suited final concentrations in the preservation mix of 1.5% (w/v) for PEG, 0.17% (w/v) for GEL and 0.07% (w/v) - 20/1 (TRH/nucleic acid w/w) - for TRH, respectively. Under these conditions PCR efficiencies did not deviate significantly from non-coated, freshly pipetted control reactions even after three months of storage at room temperature or under thermal stress at constantly 50 -¦C simulating an ageing effect. Among the three reagents TRH showed best performance since not only the most labile component - the fluorescent probe - is protected from degradation but also double-stranded plasmid DNA which can be used as reference material and in quantitative real-time PCR. -® 2011 Elsevier Ltd

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Zagon,J. / Kurth,S. / Ehlers,A. / et al: Preservation of primer and probes on "ready-to-use" 96-well microtiter plates: A step forward towards enhancing throughput and harmonization of real-time PCR applications in food and feed control. 2012.

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