Beier M, Hoheisel J D
Functional Genome Analysis, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 506, D-69120 Heidelberg, Germany.
Nucleic Acids Res. 1999 May 1;27(9):1970-7. doi: 10.1093/nar/27.9.1970.
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.
已开发出一种化学方法,该方法允许在DNA阵列上实现预制核酸(如寡核苷酸、聚合酶链反应产物或肽核酸寡聚物)的共价固定,以及在玻璃或聚丙烯表面上原位合成此类化合物。即使经过约30次洗脱循环后,发现键合仍很稳定。由于连接分子的树枝状结构,可以以可控方式改变负载量,并使其超过玻璃的容量,而不会对杂交效率产生负面影响。此外,该化学方法保证了对其他表面性质(如电荷或疏水性)的调节。核酸优先仅通过氨基修饰寡核苷酸的末端氨基或未修饰分子的末端羟基进行连接,从而使整个分子可用于探针杂交。这种衍生化代表了一种通用性足够强的支持化学方法,几乎可用于当前所有形式的DNA阵列或微芯片。