van den Berg A, van der Leij J, Poppema S
Department of Pathology and Laboratory Medicine, University Hospital Groningen, PO Box 30.001, 9700 Groningen, The Netherlands.
Nucleic Acids Res. 1999 Sep 1;27(17):e17. doi: 10.1093/nar/27.17.e17.
In a pilot study on SAGE on Reed-Sternberg cells we have sequenced 1055 tags representing 701 genes. Screening of the GenBank database resulted in the identification of a corresponding gene or EST for 490 of them. For 211 of the tags no homology could be detected. A major problem of the serial analysis of gene expression (SAGE) approach is how to further analyse the unknown tags. We have developed an RT-PCR-based method, rapid analysis of unknown SAGE tags (RAST-PCR), to analyse the expression of the corresponding genes. This approach can be used as a screening method to investigate whether or not the gene is differentially expressed between several cell types of interest.
在一项关于里德-施特恩贝格细胞的基因表达连续分析(SAGE)的初步研究中,我们对代表701个基因的1055个标签进行了测序。对GenBank数据库的筛选使得其中490个标签找到了对应的基因或EST。对于211个标签,未检测到同源性。基因表达连续分析(SAGE)方法的一个主要问题是如何进一步分析未知标签。我们开发了一种基于逆转录聚合酶链反应(RT-PCR)的方法,即未知SAGE标签快速分析(RAST-PCR),以分析相应基因的表达。这种方法可作为一种筛选方法,用于研究该基因在几种感兴趣的细胞类型之间是否存在差异表达。