A PCR primer bank for quantitative gene expression analysis

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ID: 302495
2003
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Abstract
Although gene expression profiling by microarray analysis is a useful tool for assessing global levels of transcriptional activity, variability associated with the data sets usually requires that observed differences be validated by some other method, such as real-time quantitative polymerase chain reaction (real-time PCR). However, non-specific amplification of non-target genes is frequently observed in the latter, confounding the analysis in approximately 40% of real-time PCR attempts when primer-specific labels are not used. Here we present an experimentally validated algorithm for the identification of transcript-specific PCR primers on a genomic scale that can be applied to real-time PCR with sequence-independent detection methods. An online database, PrimerBank, has been created for researchers to retrieve primer information for their genes of interest. PrimerBank currently contains 147 404 primers encompassing most known human and mouse genes. The primer design algorithm has been tested by conventional and real-time PCR for a subset of 112 primer pairs with a success rate of 98.2%.
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openalex_W2150769574 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors X. Wang
Journal Nucleic Acids Research
Year 2003
DOI
10.1093/nar/gng154
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Keywords Keywords not found

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