核酸与磁珠、Dynabeads共价结合方法的评估以及杂交反应中结合核酸的特性。
Assessment of methods for covalent binding of nucleic acids to magnetic beads, Dynabeads, and the characteristics of the bound nucleic acids in hybridization reactions.
作者信息
Lund V, Schmid R, Rickwood D, Hornes E
机构信息
Apothekernes Laboratorium A.S., Tromsø, Norway.
出版信息
Nucleic Acids Res. 1988 Nov 25;16(22):10861-80. doi: 10.1093/nar/16.22.10861.
Abstract
Dynabeads are magnetic monosized beads with high stability, high uniformity, unique paramagnetic properties, low particle-particle interaction, and high dispersibility. Different reactive groups; hydroxyl, carboxyl and amino groups can be attached to the surface. Several methods for covalent attachment of DNA or oligonucleotides to the beads were investigated. Best coupling yields were obtained by carbodiimide-mediated end-attachment of 5'-phosphate and 5'-NH2 modified nucleic acids to respectively amino and carboxyl beads. The carboxyl beads showed a low degree of non-specific binding, while a better yield of end-attached nucleic acids was obtained using the amino beads. The DNA-beads worked efficiently in hybridization experiments, and the kinetics of hybridization approach those of solution hybridization.
摘要
磁珠是具有高稳定性、高均匀性、独特顺磁性、低颗粒间相互作用和高分散性的磁性单尺寸珠子。不同的反应基团，如羟基、羧基和氨基，可以连接到其表面。研究了几种将DNA或寡核苷酸共价连接到磁珠上的方法。通过碳二亚胺介导的将5'-磷酸和5'-NH2修饰的核酸分别末端连接到氨基磁珠和羧基磁珠上，获得了最佳的偶联产率。羧基磁珠显示出较低的非特异性结合程度，而使用氨基磁珠获得了更好的末端连接核酸产率。DNA磁珠在杂交实验中有效工作，杂交动力学接近溶液杂交的动力学。
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