基于亲和的杂交捕获法对聚合酶链反应产物进行定量分析。
Quantification of polymerase chain reaction products by affinity-based hybrid collection.
作者信息
Syvänen A C, Bengtström M, Tenhunen J, Söderlund H
机构信息
Orion Genetic Engineering Laboratory, Orion Corporation Ltd, Helsinki, Finland.
出版信息
Nucleic Acids Res. 1988 Dec 9;16(23):11327-38. doi: 10.1093/nar/16.23.11327.
Abstract
We have used oligonucleotides modified with biotin in the 5'-end as primers in the polymerase chain reaction (PCR)-amplification. This results in the synthesis of 5'-biotinylated DNA molecules, which are detected by hybridization to a labelled probe in solution. The formed hybrids are collected on an avidin-matrix by mediation of the biotin residue of the target molecules. The affinity-based hybrid collection method is quantitative and makes it possible to measure the amount of DNA produced in the PCR-amplification. At low concentrations of template the efficiency of the process is close to 100%, making it possible to detect the presence of a few molecules of target DNA in 25 cycles. With high template concentrations the efficiency of the process is low.
摘要
我们使用了5'-端用生物素修饰的寡核苷酸作为聚合酶链反应（PCR）扩增的引物。这导致合成5'-生物素化的DNA分子，通过与溶液中的标记探针杂交来检测这些分子。形成的杂交体通过靶分子的生物素残基介导收集在抗生物素蛋白基质上。基于亲和力的杂交体收集方法是定量的，并且能够测量PCR扩增中产生的DNA量。在低模板浓度下，该过程的效率接近100%，使得在25个循环中能够检测到几个靶DNA分子的存在。在高模板浓度下，该过程的效率较低。
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