SEQUENCE-SPECIFIC DNA PURIFICATION BY TRIPLEX AFFINITY CAPTURE
SEQUENCE-SPECIFIC DNA PURIFICATION BY TRIPLEX AFFINITY CAPTURE
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DOI：
10.1073/pnas.89.2.495
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发表时间：
1992-01-15
影响因子：
11.1
通讯作者：
CANTOR, CR
中科院分区：
文献类型：
--
作者：
ITO, T;SMITH, CL;CANTOR, CR
利用三螺旋形成和磁分离技术建立了DNA分离方法。在该过程中，靶DNA通过分子间三链体形成被生物素化的寡核苷酸捕获，结合到链霉亲和素包被的磁珠上，并通过用使三螺旋不稳定的温和碱性缓冲液洗脱以双链形式回收。通过人工重建文库的模型实验以及（dT-dC）n的分离证明了该方法的有效性。（dG-dA）n二核苷酸重复序列。该方法为其他三链体介导的DNA分离技术提供了原型。
A DNA isolation procedure was developed by using triple-helix formation and magnetic separation. In this procedure, target DNA is captured by a biotinylated oligonucleotide via intermolecular triplex formation, bound to streptavidin-coated magnetic beads, and recovered in double-stranded form by elution with a mild alkaline buffer that destabilizes the triple helix. The effectiveness of the procedure was demonstrated by a model experiment with an artificially reconstructed library and, also, by the isolation of (dT-dC)n.(dG-dA)n dinucleotide repeats from a human genomic library. This procedure provides a prototype for other triplex-mediated DNA isolation technologies.