Lysov Iu P, Chernyĭ A A, Balaev A A, Gnuchev F N, Beattie K L, Mirzabekov A D
Mol Biol (Mosk). 1994 Jul-Aug;28(4):832-9.
The opportunity of enhancing the sequencing efficiency by applying continuous stacking hybridization is considered. The approach is based on the increase of duplex length by continuous stacking hybridization of oligonucleotides added to solution (l-oligonucleotides) to oligonucleotides immobilized on matrix (L-oligonucleotides). An analysis of reconstruction efficiency for sequenced fragments up to length of 30000 nucleotides was made. Various combinations of L- and l-oligonucleotide length were considered. The results obtained enable one to evaluate the potentialities of the proposed method for various nucleotide matrices and the complexity of experiment. Use of continuous stacking hybridization permits a considerable increase of the length of sequenced DNA fragments. We offer the approach for resolving ambiguities in branching points, which occur because of long repeats. It is based on continuous stacking hybridization of several l-oligonucleotides which form a "chain" stabilized by mutual stacking interaction.
考虑了通过应用连续堆积杂交提高测序效率的机会。该方法基于通过将添加到溶液中的寡核苷酸（l-寡核苷酸）与固定在基质上的寡核苷酸（L-寡核苷酸）进行连续堆积杂交来增加双链长度。对长度达30000个核苷酸的测序片段的重建效率进行了分析。考虑了L-和l-寡核苷酸长度的各种组合。所获得的结果使人们能够评估所提出方法对于各种核苷酸基质的潜力以及实验的复杂性。使用连续堆积杂交可显著增加测序DNA片段的长度。我们提供了解决由于长重复序列而在分支点出现的歧义的方法。它基于几种l-寡核苷酸的连续堆积杂交，这些寡核苷酸通过相互堆积相互作用形成一个“链”。
