Mies C
Department of Pathology, Jackson Memorial Hospital, Miami, Florida.
J Histochem Cytochem. 1994 Jun;42(6):811-3. doi: 10.1177/42.6.7514626.
Described here is a simple, rapid technique for isolating RNA from archived formalin-fixed, paraffin-embedded tissues (PETs). Using this modified acid guanidinium thiocyanate method, RNA sufficient as a template for the reverse transcription-polymerase chain reaction (RT-PCR) can be isolated in 2 hr from a single 20-microns-thick section of tissue. Spliced mRNA corresponding to portions of the estrogen receptor (ER) gene was successfully amplified from fixed, embedded human breast cancers containing increased amounts of ER protein. This method makes it feasible to safely and efficiently isolate RNA from large numbers of routinely processed paraffin blocks for retrospective studies of endogenous proteins such as hormone receptors, oncogenes, and viruses.
本文描述了一种从存档的福尔马林固定石蜡包埋组织（PET）中分离RNA的简单、快速技术。使用这种改良的异硫氰酸胍法，可在2小时内从20微米厚的单组织切片中分离出足以作为逆转录聚合酶链反应（RT-PCR）模板的RNA。从含有大量雌激素受体（ER）蛋白的固定、包埋的人类乳腺癌中成功扩增出与ER基因部分相对应的剪接mRNA。该方法使得从大量常规处理的石蜡块中安全、有效地分离RNA成为可能，用于对激素受体、癌基因和病毒等内源性蛋白质的回顾性研究。
