Levina Asya, Pyshnaya Inna, Repkova Marina, Rar Vera, Zarytova Valentina
Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia.
Biotechnol J. 2007 Jul;2(7):879-85. doi: 10.1002/biot.200700027.
Various materials, such as glass, plastic, metals, etc., are utilized for preparing DNA chips. In each particular case special approaches are used for immobilization of different oligonucleotide derivatives on the solid supports. We describe a general technique for DNA chips preparation on various unmodified surfaces using one type of oligonucleotide derivative, polylysine-oligonucleotide conjugates (PL-oligo). A long polyamine spacer in the PL-oligo conjugates provides a durable irreversible non-covalent immobilization onto a variety of solid supports and enough distance between oligonucleotides and the surface. The resulting DNA chips were shown to be useful for the detection of PCR DNA fragments and to be sensitive to single nucleotide discrepancies. They represent a promising instrument for revealing genetic diseases, genotyping viruses and bacteria, and for displaying their drug-resistant strains.
各种材料,如玻璃、塑料、金属等,都被用于制备DNA芯片。在每种特定情况下,都使用特殊方法将不同的寡核苷酸衍生物固定在固体支持物上。我们描述了一种使用一种寡核苷酸衍生物——聚赖氨酸-寡核苷酸缀合物(PL-寡核苷酸)在各种未修饰表面制备DNA芯片的通用技术。PL-寡核苷酸缀合物中的长聚胺间隔臂可实现对多种固体支持物的持久、不可逆非共价固定,并在寡核苷酸与表面之间提供足够的距离。结果表明,所得的DNA芯片可用于检测PCR DNA片段,并且对单核苷酸差异敏感。它们是用于揭示遗传疾病、对病毒和细菌进行基因分型以及展示其耐药菌株的一种有前景的工具。