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Nucleic Acids Research, Vol 27, Issue 18 e22-e22, Copyright © 1999 by Oxford University Press

ARTICLES

Tandem arrayed ligation of expressed sequence tags (TALEST): a new method for generating global gene expression profiles

DG Spinella, AK Bernardino, AC Redding, P Koutz, Y Wei, EK Pratt, KK Myers, G Chappell, S Gerken and SJ McConnell
Chugai Biopharmaceuticals, Inc., 6275 Nancy Ridge Drive, San Diego, CA 92121, USA. dspinella@chugaibio.com

We have developed a new and simple method for quantitatively analyzing global gene expression profiles from cells or tissues. The process, called TALEST, or tandem arrayed ligation of expressed sequence tags, employs an oligonucleotide adapter containing a type IIs restriction enzyme site to facilitate the generation of short (16 bp) ESTs of fixed position in the mRNA. These ESTs are flanked by GC-clamped punctuation sequences which render them resistant to thermal denaturation, allowing their concatenation into long arrays and subsequent recognition and analysis by high-throughput DNA sequencing. A major advantage of the TALEST technique is the avoidance of PCR in all stages of the process and hence the attendant sequence-specific amplification biases that are inherent in other gene expression profiling methods such as SAGE, Differential Display, AFLP, etc. which rely on PCR.
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