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• W2071561174 abstract "We previously reported a method for quantitative multi-gene transcriptional profiling with gene-specific standard curves using real-time PCR. Here, we report an approach that increases experimental throughput by using a master template to generate a single standard curve for the estimation of mRNA copy numbers from all genes. We prepared fifty-nine different templates and measured eNOS mRNA copy numbers in Matrigel VEGF-transfectant samples. The copy numbers obtained using each of the fifty-nine templates were within 50% of the copy number obtained using the previously reported method. Analysis of primer design parameters, and subsequent tests, showed that eliminating complementarities between the first nucleotides at the 5′-ends of the forward and reverse primers reduces the influence of saturation effects and produces copy numbers similar to those generated with gene-specific templates—generally, within 20%. Measurements on a panel consisting of eNOS, iNOS, and nNOS further validated the master-template approach. The master-template approach enables rapid quantification of mRNA abundances from panels of hundreds of genes, and will be a valuable tool for screening large numbers of genes as part of a search for biomarkers, the validation of DNA-microarray data, or research into the dynamics of the gene–protein network." @default.
• W2071561174 created "2016-06-24" @default.
• W2071561174 creator A5030726639 @default.
• W2071561174 creator A5087421256 @default.
• W2071561174 date "2005-08-01" @default.
• W2071561174 modified "2023-09-27" @default.
• W2071561174 title "Quantitative multi-gene transcriptional profiling using real-time PCR with a master template" @default.
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• W2071561174 doi "https://doi.org/10.1016/j.yexmp.2005.03.004" @default.
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