一种用于生成保留含有重复元件的克隆的消减cDNA文库的方法。
A method for generating subtractive cDNA libraries retaining clones containing repetitive elements.
作者信息
Lin C T, Sargan D R
机构信息
Department of Clinical Veterinary Medicine, Centre for Veterinary Sciences, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK.
出版信息
Nucleic Acids Res. 1997 Nov 1;25(21):4427-8. doi: 10.1093/nar/25.21.4427.
Here we describe a two-stepped photobiotin-based procedure to enrich a target (canine retinal) cDNA library for tissue specific clones without removing those containing repetitive ( SINE ) elements, despite the presence of these elements in the driver population. In a first hybridization excess SINE elements were hybridized to a driver (canine cerebellar) cDNA. In a second hybridization target cDNA was added to this reaction. The resulting cDNA library was enriched for retinal specific clones, but contained the same ratio of clones with SINE elements found in the unsubtracted library.
在此，我们描述了一种基于光生物素的两步法，用于富集目标（犬视网膜）cDNA文库以获得组织特异性克隆，而无需去除那些含有重复（短散在核元件）元件的克隆，尽管驱动群体中存在这些元件。在第一次杂交中，过量的短散在核元件与驱动（犬小脑）cDNA杂交。在第二次杂交中，将目标cDNA加入到该反应中。所得的cDNA文库富含视网膜特异性克隆，但含有与未扣除文库中发现的具有短散在核元件的克隆相同比例的克隆。
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