Nilsson M, Malmgren H, Samiotaki M, Kwiatkowski M, Chowdhary B P, Landegren U
Beijer Laboratory, Department of Medical Genetics, Biomedical Center, Uppsala, Sweden.
Science. 1994 Sep 30;265(5181):2085-8. doi: 10.1126/science.7522346.
Nucleotide sequence information derived from DNA segments of the human and other genomes is accumulating rapidly. However, it frequently proves difficult to use such short DNA segments to identify clones in genomic libraries or fragments in blots of the whole genome or for in situ analysis of chromosomes. Oligonucleotide probes, consisting of two target-complementary segments, connected by a linker sequence, were designed. Upon recognition of the specific nucleic acid molecule the ends of the probes were joined through the action of a ligase, creating circular DNA molecules catenated to the target sequence. These probes thus provide highly specific detection with minimal background.
来自人类和其他基因组DNA片段的核苷酸序列信息正在迅速积累。然而,经常证明难以使用如此短的DNA片段来鉴定基因组文库中的克隆、全基因组印迹中的片段或用于染色体的原位分析。设计了由两个与靶标互补的片段组成的寡核苷酸探针,这两个片段由一个接头序列连接。在识别特定核酸分子后,探针的末端通过连接酶的作用连接在一起,形成与靶序列连环的环状DNA分子。因此,这些探针以最小的背景提供了高度特异性的检测。