Background: Quantitative gene expression analysis by real-time PCR is important in several diagnostic areas, such as the detection of minimum residual disease in leukemia and the prognostic assessment of cancer patients. To address quality assurance in this technically challenging area, the European Union (EU) has funded the EQUAL project to develop methodologic external quality assessment (EQA) relevant to diagnostic and research laboratories among the EU member states. We report here the results of the EQUAL-quant program, which assesses standards in the use of TaqManTM probes, one of the most widely used assays in the implementation of real-time PCR. Methods: The EQUAL-quant reagent set was developed to assess the technical execution of a standard TaqMan assay, including RNA extraction, reverse transcription, and real-time PCR quantification of target DNA copy number.
EQUAL-quant: an international external quality assessment scheme for real-time PCR / Simon C Ramsden; Sarah Daly; Wolf-Jochen Geilenkeuser; Graeme Duncan; Fabienne Hermitte; Ettore Marubini; Michael Neumaier; Claudio Orlando; Vladimir Palicka; Angelo Paradiso; Mario Pazzagli; Sara Pizzamiglio; Paolo Verderio. - In: CLINICAL CHEMISTRY. - ISSN 0009-9147. - STAMPA. - 52:(2006), pp. 1584-1591.
EQUAL-quant: an international external quality assessment scheme for real-time PCR.
ORLANDO, CLAUDIO;PAZZAGLI, MARIO;
2006
Abstract
Background: Quantitative gene expression analysis by real-time PCR is important in several diagnostic areas, such as the detection of minimum residual disease in leukemia and the prognostic assessment of cancer patients. To address quality assurance in this technically challenging area, the European Union (EU) has funded the EQUAL project to develop methodologic external quality assessment (EQA) relevant to diagnostic and research laboratories among the EU member states. We report here the results of the EQUAL-quant program, which assesses standards in the use of TaqManTM probes, one of the most widely used assays in the implementation of real-time PCR. Methods: The EQUAL-quant reagent set was developed to assess the technical execution of a standard TaqMan assay, including RNA extraction, reverse transcription, and real-time PCR quantification of target DNA copy number.File | Dimensione | Formato | |
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