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- 저자 : Corbisier, Philippe (검색결과 2 건)
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White, H,Deprez, L,Corbisier, P,Hall, V,Lin, F,Mazoua, S,Trapmann, S,Aggerholm, A,Andrikovics, H,Akiki, S,Barbany, G,Boeckx, N,Bench, A,Catherwood, M,Cayuela, J-M,Chudleigh, S,Clench, T,Colomer, D,Dar Nature Publishing Group 2015 Leukemia Vol.29 No.2
<P>Serial quantification of <I>BCR–ABL1</I> mRNA is an important therapeutic indicator in chronic myeloid leukaemia, but there is a substantial variation in results reported by different laboratories. To improve comparability, an internationally accepted plasmid certified reference material (CRM) was developed according to ISO Guide 34:2009. Fragments of <I>BCR–ABL1</I> (e14a2 mRNA fusion)<I>, BCR</I> and <I>GUSB</I> transcripts were amplified and cloned into pUC18 to yield plasmid pIRMM0099. Six different linearised plasmid solutions were produced with the following copy number concentrations, assigned by digital PCR, and expanded uncertainties: 1.08±0.13 × 10<SUP>6</SUP>, 1.08±0.11 × 10<SUP>5</SUP>, 1.03±0.10 × 10<SUP>4</SUP>, 1.02±0.09 × 10<SUP>3</SUP>, 1.04±0.10 × 10<SUP>2</SUP> and 10.0±1.5 copies/μl. The certification of the material for the number of specific DNA fragments per plasmid, copy number concentration of the plasmid solutions and the assessment of inter-unit heterogeneity and stability were performed according to ISO Guide 35:2006. Two suitability studies performed by 63 <I>BCR–ABL1</I> testing laboratories demonstrated that this set of 6 plasmid CRMs can help to standardise a number of measured transcripts of e14a2 <I>BCR–ABL1</I> and three control genes (<I>ABL1, BCR</I> and <I>GUSB</I>). The set of six plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (https://ec.europa.eu/jrc/en/reference-materials/catalogue/; CRM code ERM-AD623a-f).</P>
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Yoo, Hee-Bong,Park, Sang-Ryoul,Dong, Lianhua,Wang, Jing,Sui, Zhiwei,Pavš,ič,, Jernej,Milavec, Mojca,Akgoz, Muslum,Mozioğ,lu, Erkan,Corbisier, Philippe,Janka, Má,trai,Cosme, Bru American Chemical Society 2016 ANALYTICAL CHEMISTRY - Vol.88 No.24
<P>Enumeration-based determination of DNA copy-concentration was assessed through an international comparison among national metrology institutes (NMIs) and designated institutes (DIs). Enumeration-based quantification does not require a calibration standard thereby providing a route to 'absolute quantification', which offers the potential for reliable value assignments of DNA reference materials, and International System of Units (SI) traceability to copy number 1 through accurate counting. In this study, 2 enumeration based methods, flow cytometric (FCM) counting and the digital polymerase chain reaction (dPCR), were compared to quantify a solution of the pBR322 plasmid at a concentration of several thousand copies per microliter. In addition, 2 orthogonal chemical-analysis methods based on nucleotide quantification, isotope-dilution mass spectrometry (IDMS) and capillary electrophoresis (CE) were applied to quantify a more concentrated solution of the plasmid. Although 9 dPCR results from 8 laboratories showed some dispersion (relative standard deviation [RSD] = 11.8%), their means were closely aligned with those of the FCM-based counting method and the orthogonal chemical-analysis methods, corrected for gravimetric dilution factors. Using the means of dPCR results, the RSD of all 4 methods was 1.8%, which strongly supported the validity of the recent enumeration approaches. Despite a good overall agreement, the individual dPCR results were not sufficiently covered by the reported measurement uncertainties. These findings suggest that some laboratories may not have considered all factors contributing to the measurement uncertainty of dPCR, and further investigation of this possibility is warranted.</P>
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