摘要
以Ｎ－（４－马来酰亚胺氧化丁酰）－琥珀酰亚胺磺酸钠盐（ｓｕｌｆｏ－ＧＭＢＳ）为偶联剂将５’－氨基／３’－得克萨斯红双末端修饰ＤＮＡ探针偶联到３－（２－氨乙基氨丙基）三甲氧基硅烷（ＡＡＴＳ）修饰的３００μｍＺｒＯ２陶瓷小珠表面，制成荧光小珠。小珠表面上单链ＤＮＡ探针分子之间的距离取决于小珠表面的预处理、偶联反应溶液中ＤＮＡ探针浓度、小珠大小和小珠的数目。在优化条件下，２０－ｍｅｒ单链ＤＮＡ探针在小珠表面微阵列的最小分子间距约为７．６ｎｍ，即在一个３００μｍＺｒＯ２陶瓷小珠表面有１０１０数量级的２０－ｍｅｒ单链 ＤＮＡ探针参与微阵列反应。机理研究表明：ＤＮＡ探针通过ｓｕｌｆｏＧＧＭＢＳ偶联到小珠表面的化学反应仅涉及到探针末端修饰氨基。
The microarray features of 5'- NH2/3'- Texas red terminus modified deoxyribonucleic acid (DNA) probes on the surfaces of 300 mum ZrO2 ceramic beads were characterized through the coupling reaction of the probes with N-(4-maleimidobutyryloxy) sulfosuccininimide sodium salt (sulfa-GMBS). Based on the fluorescence measurement on the bead surface, it was found that the microarray of DNA probes depends on the pretreatment of the bead surface, DNA probe concentrations in aqueous solution and the total surface area of the beads. On optimal conditions, a 20-mer DNA probe can dot on the beads surface with the molecule distance of 7.6 nm, and more than 1.0 x 10(10) 20-mer single-stranded DNA probe can be microarrayed on one ZrO2 bead surface. Mechanism study shows that the coupling reaction of DNA probe on the ceramic bead surface only involves in the terminus NH2-group.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2002年第1期13-17,共5页
Chinese Journal of Analytical Chemistry
基金
教育部高等学校骨干教师资助计划(文号:教技司2000-65)
国家自然科学基金(No.20045002)
重庆市科委应用基础
