Southern E M, Maskos U, Elder J K
Department of Biochemistry, University of Oxford, United Kingdom.
Genomics. 1992 Aug;13(4):1008-17. doi: 10.1016/0888-7543(92)90014-j.
An efficient method was developed for making complete sets of oligonucleotides of defined length, covalently attached to the surface of a glass plate, by synthesizing them in situ. A device carrying all octapurine sequences was used to explore factors affecting molecular hybridization of the tethered oligonucleotides, to develop computer-aided methods for analyzing the data, and to test the feasibility of using the method for sequence analysis. Further development is needed before the method can be used routinely, but our work shows that it has a number of potential advantages over gel-based methods: it should be easy to automate; the quality of the sequence results can be evaluated statistically; it provides a powerful way of comparing related sequences and detecting mutation; it can be applied to both DNA and RNA; and specific motifs can be incorporated into all sequences of the array to focus analysis on sequences of biological interest.
通过原位合成，开发出一种高效方法，可制备共价连接在玻璃板表面的特定长度的寡核苷酸完整集。使用携带所有八聚嘌呤序列的装置来探索影响固定化寡核苷酸分子杂交的因素，开发用于分析数据的计算机辅助方法，并测试使用该方法进行序列分析的可行性。在该方法能够常规使用之前还需要进一步改进，但我们的工作表明，它相对于基于凝胶的方法具有许多潜在优势：易于自动化；序列结果的质量可以进行统计学评估；它提供了一种比较相关序列和检测突变的强大方法；它可应用于DNA和RNA；并且可以将特定基序纳入阵列的所有序列中，以便将分析集中在具有生物学意义的序列上。
