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(American Journal of Pathology. 2004;165:1799-1807.)
© 2004 American Society for Investigative Pathology

Quantitative Gene Expression Profiling in Formalin-Fixed, Paraffin-Embedded Tissues Using Universal Bead Arrays

Marina Bibikova*, Dimitri Talantov{dagger}, Eugene Chudin*, Joanne M. Yeakley*, Jing Chen*, Dennis Doucet*, Eliza Wickham*, David Atkins{dagger}, David Barker*, Mark Chee*, Yixin Wang{dagger} and Jian-Bing Fan*

From Illumina, Inc.,* San Diego; and Veridex, LLC,{dagger} a Johnson & Johnson Company, San Diego, California

We recently developed a sensitive and flexible gene expression profiling system that is not dependent on an intact poly-A tail and showed that it could be used to analyze degraded RNA samples. We hypothesized that the DASL (cDNA-mediated annealing, selection, extension and ligation) assay might be suitable for the analysis of formalin-fixed, paraffin-embedded tissues, an important source of archival tissue material. We now show that, using the DASL assay system, highly reproducible tissue- and cancer-specific gene expression profiles can be obtained with as little as 50 ng of total RNA isolated from formalin-fixed tissues that had been stored from 1 to over 10 years. Further, tissue- and cancer-specific markers derived from previous genome-wide expression profiling studies of fresh-frozen samples were validated in the formalin-fixed samples. The DASL assay system should prove useful for high-throughput expression profiling of archived clinical samples.




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