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Quantitative gene expression profiling in formalin-fixed, paraffin-embedded tissues using universal bead arrays.


ABSTRACT: 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.

SUBMITTER: Bibikova M 

PROVIDER: S-EPMC1618650 | biostudies-literature | 2004 Nov

REPOSITORIES: biostudies-literature

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Quantitative gene expression profiling in formalin-fixed, paraffin-embedded tissues using universal bead arrays.

Bibikova Marina M   Talantov Dimitri D   Chudin Eugene E   Yeakley Joanne M JM   Chen Jing J   Doucet Dennis D   Wickham Eliza E   Atkins David D   Barker David D   Chee Mark M   Wang Yixin Y   Fan Jian-Bing JB  

The American journal of pathology 20041101 5


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 c  ...[more]

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