Chetverin A B, Kramer F R
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region.
Biotechnology (N Y). 1994 Nov;12(11):1093-9. doi: 10.1038/nbt1194-1093.
Advances in solid-phase oligonucleotide synthesis and hybridization techniques have led to an incipient technology based on the use of oligonucleotide arrays. The inclusion of a large number of oligonucleotide probes within a single array greatly reduces the cost of their synthesis and allows thousands of hybridizations to be carried out simultaneously. The range of potential applications of oligonucleotide arrays was expanded by the realization that nucleic acids can be sequenced by hybridizing them to all possible oligonucleotides of a given length. Additional possibilities are offered by novel types of oligonucleotide arrays that are capable of parallel sorting, isolating, and manipulating thousands, and even millions, of nucleic acid species. Fields, such as site-directed mutagenesis, protein engineering, and recombinant DNA technology, would benefit from using these arrays. Further, these approaches could enable the analysis of entire genomes by preparing ordered fragment libraries, and by sequencing complex pools of nucleic acids, in a novel approach that provides long-range sequence information by generating nested nucleic acids and then surveying the oligonucleotides contained in the nested strands. This would allow large diploid genomes to be sequenced directly in a completely automated procedure that does not require fragment cloning or chromosome mapping.
固相寡核苷酸合成和杂交技术的进步催生了一种基于寡核苷酸阵列使用的新兴技术。在单个阵列中包含大量寡核苷酸探针极大地降低了其合成成本，并允许同时进行数千次杂交。意识到核酸可以通过与给定长度的所有可能寡核苷酸杂交来测序，寡核苷酸阵列的潜在应用范围得到了扩展。新型寡核苷酸阵列能够对数千甚至数百万种核酸进行平行分选、分离和操作，提供了更多可能性。定点诱变、蛋白质工程和重组DNA技术等领域将受益于使用这些阵列。此外，这些方法可以通过制备有序片段文库，并以一种通过生成嵌套核酸然后检测嵌套链中包含的寡核苷酸来提供长程序列信息的新方法对复杂的核酸池进行测序，从而实现对整个基因组的分析。这将允许以完全自动化的程序直接对大型二倍体基因组进行测序，而无需片段克隆或染色体作图。
