Quantitative gene expression profiling in formalin-fixed, paraffin-embedded tissues using universal bead arrays
Quantitative gene expression profiling in formalin-fixed, paraffin-embedded tissues using universal bead arrays
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DOI：
10.1016/s0002-9440(10)63435-9
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发表时间：
2004-11-01
影响因子：
6
通讯作者：
Fan, JB
中科院分区：
文献类型：
--
作者：
Bibikova, M;Talantov, D;Fan, JB
我们最近开发了一种灵敏而灵活的基因表达谱系统，它不依赖于完整的Poly-A尾巴，并表明它可以用于分析降解的RNA样本。我们推测，DASL(DNA介导的退火、选择、延伸和连接)方法可能适用于福尔马林固定的石蜡包埋组织的分析，而福尔马林固定的石蜡包埋组织是档案组织材料的重要来源。我们现在表明，使用DASL分析系统，从福尔马林固定的组织中提取的总RNA只要50 ng，就可以获得高度重复性的组织和癌症特异性基因表达谱，这些组织已经储存了1到10年以上。此外，从先前对新鲜冷冻样本的全基因组表达谱研究中获得的组织和癌症特异性标记在福尔马林固定的样本中得到了验证。DASL分析系统应该被证明对存档的临床样本的高通量表达谱是有用的。
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 I 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.