DNA SEQUENCING - MODULAR PRIMERS ASSEMBLED FROM A LIBRARY OF HEXAMERS OR PENTAMERS
DNA SEQUENCING - MODULAR PRIMERS ASSEMBLED FROM A LIBRARY OF HEXAMERS OR PENTAMERS
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DOI：
10.1073/pnas.90.9.4241
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发表时间：
1993-05-01
影响因子：
11.1
通讯作者：
ULANOVSKY, LE
中科院分区：
文献类型：
--
作者：
KOTLER, LE;ZEVINSONKIN, D;ULANOVSKY, LE
在这里，我们报告了一种显著的效果，它由六聚体或五聚体寡核苷酸模块组成，在退火到DNA模板后彼此碱基堆叠。这种模块的组合被发现可以独特地启动DNA测序反应，而不是单独使用任何一个模块。我们将这种效应部分归因于在存在相邻模块的情况下，寡核苷酸对模板的亲和力增加。所有可能的五聚体(或六聚体)序列总计1024个(或4096个)样本，对于预先合成的文库来说，这是一个可管理的大小。这种方法可以取代目前在时间和成本上的瓶颈--引物的合成，并且可以实现闭合循环行走的全自动化。
Here we report a striking effect displayed by ''modular primers,'' which consist of hexamer or pentamer oligonucleotide modules base-stacked to each other upon annealing to a DNA template. Such a combination of modules is found to prime DNA sequencing reactions uniquely, unlike either of the modules alone. We attribute this effect in part to the increase in the affinity of an oligonucleotide for the template in the presence of an adjacent module. All possible pentamer (or hexamer) sequences total 1024 (or 4096) samples, a manageable size for a presynthesized library. This approach can replace the synthesis of primers, which is the current bottleneck in time and cost of the primer walking sequencing, and can allow full automation of the closed cycle of walking.