Taylor G P, Troyer D A, Giambernardi T A, Klebe R J
Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio 78284, USA.
J Pathol. 1998 Mar;184(3):332-5. doi: 10.1002/(SICI)1096-9896(199803)184:3<332::AID-PATH985>3.0.CO;2-U.
The sensitivity of the reverse transcriptase-polymerase chain reaction (RT-PCR) makes it ideally suited for the detection of changes in gene expression. Unfortunately, traditional methods for RNA isolation require time-consuming procedures that are not appropriate for small samples, such as individual frozen sections. This report describes a new technique that permits the rapid extraction of RNA from individual frozen histological sections. RNA is extracted by incubating a frozen section in an RT-PCR compatible buffer solution containing RNase inhibitor and dithiothreitol. RNA isolated from frozen sections is stable at room temperature for up to 3 h under the conditions described. Alternatively, extracts can be frozen for later use. When maintained in a dry state at room temperature, RNA in sections remained stable for 2 weeks. Histological, immunohistochemical, or in situ analyses can be carried out with sections that are immediately adjacent to those used for extracting RNA. The simplicity and economy of this procedure may foster the development of prognostic screens that can be performed in parallel with traditional histopathological analysis.
逆转录-聚合酶链反应（RT-PCR）的高灵敏度使其非常适合用于检测基因表达的变化。不幸的是，传统的RNA分离方法需要耗时的步骤，不适用于小样本，如单个冷冻切片。本报告描述了一种新技术，可从单个冷冻组织学切片中快速提取RNA。通过将冷冻切片在含有RNase抑制剂和二硫苏糖醇的RT-PCR兼容缓冲溶液中孵育来提取RNA。在所述条件下，从冷冻切片中分离的RNA在室温下最多可稳定3小时。或者，提取物可以冷冻以备后用。当在室温下保持干燥状态时，切片中的RNA可稳定2周。可以对紧邻用于提取RNA的切片进行组织学、免疫组织化学或原位分析。该方法的简单性和经济性可能会促进与传统组织病理学分析并行进行的预后筛查的发展。
