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首页> 外文期刊>Food Control >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.
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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.

机译：在“即用型” 96孔微量滴定板上保存引物和探针：朝着提高通量和协调实时PCR在食品和饲料控制中的应用迈出了一步。

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摘要

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 (TRHucleic 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 degrees 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. All rights reserved, Elsevier.
机译：研究了三种不同的试剂海藻糖（TRH），明胶（GEL）和聚乙二醇（PEG）的保存寡核苷酸的能力，例如在96孔微量滴定板中预涂的引物和探针或双链质粒DNA。对选定的多拷贝和两个单拷贝PCR（聚合酶链反应）系统的保留效率进行评估后，发现保存混合物中最适合的最终浓度是PEG为1.5％（w / v），0.17％（w / v） ）（对于GEL）和0.07％（w / v）-20/1（TRH /核酸w / w）-对于TRH。在这些条件下，即使在室温下或在持续50摄氏度的热应力下模拟老化效果三个月后，PCR效率也不会明显偏离未涂布的新鲜移液对照反应。在这三种试剂中，TRH表现出最佳性能，因为不仅保护了最不稳定的成分（荧光探针）免于降解，而且保护了双链质粒DNA（可用作参考材料和实时定量PCR）。保留所有权利，Elsevier。

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