Use of a fluorescent-PCR reaction to detect genomic sequence copy number and transcriptional abundance

Pei Wen Chiang, Woo Joo Song, Kai Yuan Wu, Julie R. Korenberg, Eric J. Fogel, Margaret L. Van Keuren, Deval Lashkari, David M. Kurnit

Research output: Contribution to journalArticle

51 Scopus citations

Abstract

We present a fluorescent-PCR-based technique to assay genomic sequence copy number and transcriptional abundance. This technique relies on the ability to follow fluorescent PCR progressively in real time during the exponential phase of the reaction so that quantitative PCR is accomplished. We demonstrated the ability of this technique to quantitate both known deletions and amplifications of loci that have been measured previously by other methods, and to measure transcriptional abundance. Using an efficient variant of the fluorescent-PCR technology, we can monitor transcription semiquantitatively. The ability to detect all amplifications and deletions at any single copy locus by PCR makes this the technique of choice to assay genomic sequence copy number anomalies in birth defects and cancers. The ability to detect variations in transcript abundance enables this technique to fashion a time and tissue analysis of transcription.

Original languageEnglish (US)
Pages (from-to)1013-1026
Number of pages14
JournalGenome Research
Volume6
Issue number10
DOIs
StatePublished - 1996

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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    Chiang, P. W., Song, W. J., Wu, K. Y., Korenberg, J. R., Fogel, E. J., Van Keuren, M. L., Lashkari, D., & Kurnit, D. M. (1996). Use of a fluorescent-PCR reaction to detect genomic sequence copy number and transcriptional abundance. Genome Research, 6(10), 1013-1026. https://doi.org/10.1101/gr.6.10.1013