von Ahlfen Silke, Missel Andreas, Bendrat Klaus, Schlumpberger Martin
QIAGEN GmbH, Hilden, Germany.
PLoS One. 2007 Dec 5;2(12):e1261. doi: 10.1371/journal.pone.0001261.
The large archives of formalin-fixed paraffin-embedded (FFPE) tissue specimens that exist are a highly valuable source of sample material for molecular biological analysis, including gene expression profiling. However, current data on adverse effects of standard pathological practice on the usefulness of biomolecular analytes obtained from such archived specimens is largely anecdotal. Here, we present a systematic examination of the most relevant parameters for integrity and useability of RNA obtained from FFPE samples, including storage time and conditions, fixation time, and specimen size. The results are particularly relevant for any application relying on cDNA synthesis as an initial step of the procedure, such as RT-PCR, and microarray analysis.
现有的大量福尔马林固定石蜡包埋(FFPE)组织标本档案是用于分子生物学分析(包括基因表达谱分析)的极有价值的样本材料来源。然而,目前关于标准病理操作对从此类存档标本中获取的生物分子分析物有用性的不良影响的数据大多是传闻性的。在此,我们对从FFPE样本中获得的RNA完整性和可用性的最相关参数进行了系统研究,包括储存时间和条件、固定时间以及标本大小。这些结果对于任何依赖cDNA合成作为该程序初始步骤的应用(如RT-PCR和微阵列分析)尤为重要。