Mirzabekov A D
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow.
Trends Biotechnol. 1994 Jan;12(1):27-32. doi: 10.1016/0167-7799(94)90008-6.
The length of DNA from different sources that has been sequenced has already exceeded 1 x 10(8) bases, and this figure is continuing to grow exponentially. Even so, the very large quantity of DNA that remains to be sequenced and the impetus generated by the worldwide Human Genome Project has increased the need for the development of megasequencing procedures. Sequencing by hybridization (SbH) shows promise as an approach for developing such a method. The technique involves hybridization of the DNA of unknown sequence with an enormous set of short oligonucleotides; identification and analysis of the overlapping set of oligomers that form perfect duplexes with the DNA of interest permits reconstruction of the target-DNA sequence. Preliminary experiments have already demonstrated the feasibility of incorporating this approach into large-scale sequencing projects, and processes have been developed both for manufacturing the sequencing microchips that incorporate the immobilized oligonucleotides, and for detecting hybridization of the target DNA to these microchips.
目前已测序的来自不同来源的DNA长度已超过1×10⁸个碱基，而且这一数字还在呈指数级持续增长。即便如此，仍有大量DNA有待测序，再加上全球人类基因组计划所产生的推动力，对大规模测序程序的需求与日俱增。杂交测序法（SbH）有望成为开发此类方法的途径。该技术涉及将未知序列的DNA与大量短寡核苷酸进行杂交；对与目标DNA形成完美双链体的重叠寡聚体集合进行鉴定和分析，从而能够重建目标DNA序列。初步实验已证明将此方法纳入大规模测序项目的可行性，并且已开发出用于制造包含固定化寡核苷酸的测序微芯片以及检测目标DNA与这些微芯片杂交的方法。
