Guerasimova A, Ivanov I, Lehrach H
Max-Planck-Institute für Molekular Genetik, Ihnestrasse 73, 14195 Berlin-Dahlem, Germany andW. Engelhardt Institute of Molecular Biology, Vavilov strasse 34, 117984 Moscow, Russia.
Nucleic Acids Res. 1999 Jan 15;27(2):703-5. doi: 10.1093/nar/27.2.703.
Here we describe a method of labelling short oligonucleotide probes with enzyme without purification or chemical modifications. Biotinylated oligonucleotides as short as 10 nt are coupled with streptavidin-conjugated enzyme, hybridised and detected with enzyme-triggered chemiluminescence. The detection of hybridisation signal is linear for two orders of magnitude of target dilution. It is shown to be comparable in sensitivity with standard procedures and with radioactive detection. The method is quick, simple and has potential for automation of large-scale oligo-nucleotide hybridisation and multiplex sequencing.
在此，我们描述了一种无需纯化或化学修饰即可用酶标记短寡核苷酸探针的方法。短至10个核苷酸的生物素化寡核苷酸与链霉亲和素偶联酶结合，进行杂交并用酶触发的化学发光进行检测。杂交信号的检测对于两个数量级的靶标稀释呈线性关系。结果表明，其灵敏度与标准方法及放射性检测相当。该方法快速、简单，具有大规模寡核苷酸杂交和多重测序自动化的潜力。
