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首页> 外文期刊>Diagnostic molecular pathology : >Amplification of mRNA from laser-microdissected single or clustered cells in formalin-fixed and paraffin-embedded tissues for application in quantitative real-time PCR.
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Amplification of mRNA from laser-microdissected single or clustered cells in formalin-fixed and paraffin-embedded tissues for application in quantitative real-time PCR.

机译：在福尔马林固定和石蜡包埋的组织中，通过激光显微切割的单个或簇状细胞扩增mRNA，用于定量实时PCR。

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摘要

The determination of marker genes and gene clusters involved in disease pathogenesis is increasingly contingent on high-throughput methods of gene expression profiling. However, the concurrently increasing application of mRNA from formalin-fixed and paraffin-embedded (FFPE) tissue archives, as well as cell-type-specific approaches by laser-assisted microdissection, frequently results in very small and degraded quantities of RNA. Therefore, a successful amplification of cell-type-specific mRNA targets from FFPE tissues becomes more and more essential. To optimize the hitherto limited technical options, we applied 3 commercial amplification kits on FFPE single cells. We thereby determined the approach of target-specific cDNA amplification as being notably appropriate for subsequent real-time polymerase chain reaction, as a constant decrease of CT values by 14 polymerase chain reaction cycles could be demonstrated.
机译：疾病发病机理中涉及的标记基因和基因簇的确定越来越取决于基因表达谱分析的高通量方法。但是，同时增加的福尔马林固定和石蜡包埋（FFPE）组织档案中的mRNA的应用，以及通过激光辅助显微解剖的细胞类型特异性方法，通常会导致非常少量且降解的RNA。因此，从FFPE组织成功扩增细胞类型特异性mRNA靶标变得越来越重要。为了优化迄今为止有限的技术选择，我们在FFPE单细胞上应用了3种商业扩增试剂盒。因此，我们确定了目标特异性cDNA扩增的方法特别适合于随后的实时聚合酶链反应，因为可以证明14个聚合酶链反应循环会导致CT值不断降低。

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