A simple method for RNA isolation from formalin-fixed and paraffin-embedded lymphatic tissues
A simple method for RNA isolation from formalin-fixed and paraffin-embedded lymphatic tissues
复制标题
DOI：
10.1016/s0014-4800(03)00024-8
复制
发表时间：
2003-06-01
影响因子：
3.6
通讯作者：
Antica, M
中科院分区：
文献类型：
--
作者：
Körbler, T;Grskovic, M;Antica, M
淋巴细胞分化下的基因激活是近年来免疫学研究最多的问题之一。然而，对淋巴增生性疾病中这一分子事件的分析常常因缺乏新鲜材料而受到阻碍。可用于常规组织学检查的大多数组织是福尔马林固定和石蜡包埋的。这些标本中的基因表达可以通过mRNA的逆转录和聚合酶链反应（RT-PCR）进行分析。因此，我们调整并建立了一种从这些标本中分离mRNA的方法，该方法结合了先前报道的方案和对苯酚/氯仿提取方法的修改。鉴于转录因子在人造血系统中的重要作用，我们研究了能否从档案组织中成功分离mRNA，用于研究Ikaros家族转录因子在淋巴组织中的表达。虽然从档案组织中分离的RNA的定量分析可能不可行，因为不同样本的RNA分离程度不可预测，但我们在这里表明筛选分析是可能的和简单的。(C) 2003 Elsevier Science（美国）版权所有。
Gene activation that lies beneath lymphoid cell differentiation has been one of the most explored issues in immunology in the recent years. However, the analysis of this molecular event in lymphoproliferative diseases is often hampered by the lack of fresh material. Most tissues available for routine histological investigation are formalin fixed and paraffin embedded. Gene expression in such specimens could be analyzed using reverse transcription of mRNA and the polymerase chain reaction (RT-PCR). Therefore we adjusted and established a method for mRNA isolation from such specimens by a combination of previously reported protocols and a modification of the phenol/chloroform extraction method. Given the significance of transcription factors in the human hemopoietic system, we investigated whether mRNA could be successfully isolated from archival tissue for a study on expression of Ikaros family transcription factors in lymphatic tissue. Although quantitative analysis of RNA isolated from archival tissue is probably not feasible due to the unpredictable degree of RNA isolation varying from sample to sample, we show here that screening analysis is possible and simple. (C) 2003 Elsevier Science (USA). All rights reserved.