增强型串联体克隆——对基因表达序列分析（SAGE）技术的一种改进。
Enhanced concatemer cloning-a modification to the SAGE (Serial Analysis of Gene Expression) technique.
作者信息
Powell J
机构信息
The Richard Dimbleby Department of Cancer Research, I.C.R.F. Laboratory, Rayne Institute, 4th Floor Lambeth Wing, St Thomas's Hospital, Lambeth Palace Road, London SE1 7EH, UK.
出版信息
Nucleic Acids Res. 1998 Jul 15;26(14):3445-6. doi: 10.1093/nar/26.14.3445.
Abstract
The Serial Analysis of Gene Expression (SAGE) method, described in 1995 by Velculescu et al ., represents a powerful means to compare gene expression between two mRNA populations. An improvement to SAGE that removes contaminating linker molecules, which compromise the efficiency of the method, has been developed. This modification utilises biotinylated PCR primers, which generate biotinylated linkers at an early stage in the SAGE protocol, thus allowing removal of the unwanted linkers by binding to streptavidin-coated magnetic beads at a later stage. The application of this modification resulted in the rapid generation of high ditag yields and clones with large average insert sizes.
摘要
1995年，韦尔库列斯库等人描述的基因表达序列分析（SAGE）方法，是比较两个mRNA群体之间基因表达的有力手段。人们已经开发出一种对SAGE的改进方法，该方法可以去除会影响该方法效率的污染性接头分子。这种改进利用了生物素化的PCR引物，这些引物在SAGE方案的早期阶段产生生物素化接头，从而允许在后期通过与链霉亲和素包被的磁珠结合来去除不需要的接头。这种改进的应用导致了高双标签产量的快速产生以及具有大平均插入片段大小的克隆。
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