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优化在 1 乙基 3 ( 3 二甲基氨丙基 ) 碳化二亚胺 (EDC)存在下 ,5′ NH2 单末端和 5′ TR 3′ NH2 双末端修饰单链脱氧核糖核酸 (DNA)探针在 10 μm羧基聚苯乙烯小珠表面进行微阵列的条件 ,研究 5′ NH2 单末端修饰DNA探针与其碱基互补序列进行的固 液杂交特性。结果表明 ,在pH 4 .6～ 5.6,EDC浓度为 0 .5～ 0 .7g L时 ,DNA探针在小珠表面具有最大的微阵列能力。微阵列在小珠表面的 5′ NH2 单末端修饰DNA探针 ,与其碱基互补序列的杂交反应遵守二级反应动力学 ,杂交优化条件除取决于溶液介质的pH值、离子强度和碱基互补序列在溶液介质中的浓度外 ,还取决于环境温度和杂交时间。测定了 2 5℃下微阵列在小珠表面的 2 0 merDNA探针与其溶液中互补序列的杂交反应常数。
Optimization of 5 ’NH2 single-terminal and 5’ TR 3 ’NH2 dual-terminal modified single-stranded DNA probes in the presence of 1 ethyl 3 (3 dimethylaminopropyl) carbodiimide (EDC) The conditions of the microarray on the surface of 10 μm carboxyl polystyrene beads were investigated, and the solid-liquid hybridization characteristics of the 5 ’NH2 single-terminal modified DNA probe and its base complementary sequence were studied. The results showed that the DNA probe has the largest microarray ability on the surface of beads at pH 4.6 ~ 5.6 and EDC concentration of 0.5 ~ 0.7 g L. Microarray on the surface of the bead 5 ’NH2 single-ended modified DNA probes hybridization reaction with its base complementary to the second-order reaction kinetics, hybridization optimization conditions except depending on the solution medium pH, ionic strength and base complement The concentration of the sequence in the solution medium also depends on the ambient temperature and the time of hybridization. The hybridization reaction constants of 2 0 mer DNA probes of the microarray on the bead surface with the complementary sequences in the solution at 25 ° C. were measured.