首页> 美国卫生研究院文献>Nucleic Acids Research >Versatile derivatisation of solid support media for covalent bonding on DNA-microchips.
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Versatile derivatisation of solid support media for covalent bonding on DNA-microchips.

机译:固相支持介质的多功能衍生化用于在DNA微芯片上共价键合。

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摘要

A chemistry was developed that permits on DNA-arrays both the covalent immobilisation of pre-fabricated nucleic acids-such as oligonucleotides, PCR-products or peptide nucleic acid oligomers-and the in situ synthesis of such compounds on either glass or polypropylene surfaces. Bonding was found to be stable even after some 30 cycles of stripping. Due to a dendrimeric structure of the linker molecule, the loading can be modified in a controlled manner and increased beyond the capacity of glass without negative effects on hybridisation efficiency. Also, the chemistry warrants the modulation of other surface properties such as charge or hydrophobicity. Preferentially, attachment of nucleic acids takes place only via the terminal amino-group of amino-modified oligonucleotides or the terminal hydroxyl-group of unmodified molecules so that the entire molecule is accessible to probe hybridisation. This derivatisation represents a support chemistry versatile enough to serve nearly all current forms of DNA-arrays or microchips.
机译:已开发出一种化学方法,该化学方法可将预制核酸(如寡核苷酸,PCR产物或肽核酸低聚物)共价固定在DNA阵列上,并在玻璃或聚丙烯表面上原位合成此类化合物。发现即使在剥离的约30个循环后,粘合也是稳定的。由于接头分子的树枝状结构,可以以可控的方式改变载量并增加其载量,使其超过玻璃的容量,而对杂交效率没有负面影响。同样,化学性质保证其他表面性质的调节,例如电荷或疏水性。优选地,核酸的附着仅通过氨基修饰的寡核苷酸的末端氨基或未修饰分子的末端羟基发生,使得整个分子可用于探针杂交。这种衍生化反应是一种支持化学,其用途广泛,足以用于几乎所有当前形式的DNA阵列或微芯片。

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