Irving J A, Cain G, Parr A, Howard M, Angus B, Cattan A R
Department of Haematology, Royal Victoria Infirmary, Newcastle upon Tyne.
J Clin Pathol. 1996 Mar;49(3):258-9. doi: 10.1136/jcp.49.3.258.
OCT embedded cryostat sections of stored pathological specimens of non-Hodgkin's lymphoma were used to provide RNA. After reverse transcription to produce cDNA, the polymerase chain reaction was performed with primers for standard and variant forms of the CD44 molecule. Using Southern transfer and hybridisation with a probe specific for exon 4 of the CD44 gene, both standard and variant forms were visualised by autoradiography. This method was shown to be applicable to other gene products by using primers specific for the abl and bcr genes. This technique permits retrospective analysis of RNA from small amounts of stored pathological samples.
使用储存的非霍奇金淋巴瘤病理标本的OCT包埋冰冻切片来提供RNA。逆转录产生cDNA后,用针对CD44分子标准型和变异型的引物进行聚合酶链反应。通过Southern印迹转移并用针对CD44基因第4外显子的特异性探针进行杂交,通过放射自显影观察到标准型和变异型。通过使用针对abl和bcr基因的特异性引物,该方法被证明适用于其他基因产物。该技术允许对来自少量储存病理样本的RNA进行回顾性分析。