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Nucleic Acids Research, Vol 27, Issue 23 4585-4589, Copyright © 1999 by Oxford University Press

ARTICLES

In vivo analysis of cancerous gene expression by RNA-polymerase chain reaction

SL Lin, CM Chuong, RB Widelitz and SY Ying
Department of Pathology, University of Southern California School of Medicine, HMR-209, 2011 Zonal Avenue, Los Angeles, CA 90033, USA. shilungl@hsc.usc.edu

An easy and routine procedure to amplify messenger RNA (mRNA) libraries from a few tissue cells can provide molecular gene expression profiles at high resolution. A novel PCR-like method, the RNA-PCR, was developed to generate high quality and quantity mRNAs from as few as 20 cells (2 pg mRNAs). The principle relies upon the cycling steps of promoter- linked double-stranded cDNA synthesis and promoter-driven transcription to amplify mRNAs up to 250-fold/cycle with good representation of high and low copy mRNAs. The amplified mRNA libraries were shown to possess high fidelity, purity, specificity and reproducibility for in vivo analyses of cancerous gene expression in human prostate cancers.
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