Versatile derivatisation of solid support media for covalent bonding on DNA-microchips
Versatile derivatisation of solid support media for covalent bonding on DNA-microchips
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DOI：
10.1093/nar/27.9.1970
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发表时间：
1999-05-01
影响因子：
14.9
通讯作者：
Hoheisel, JD
中科院分区：
文献类型：
--
作者：
Beier, M;Hoheisel, JD
一种化学方法被开发出来，允许在dna阵列上对预制核酸（如寡核苷酸、聚合酶链反应产物或肽核酸寡聚物）进行共价固定，并在玻璃或聚丙烯表面上原位合成这些化合物。发现即使在剥离30次循环后，键合仍然稳定。由于连接分子的树突结构，负载可以以可控的方式进行修改，并且可以超过玻璃的容量而不会对杂化效率产生负面影响。此外，化学性质保证了其他表面性质的调制，如电荷或疏水性。优先地，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进行，因此整个分子都可以进行探针杂交。这种衍生化代表了一种多用途的支持化学，足以服务于几乎所有当前形式的dna阵列或微芯片。
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.