6 Base PCR to Build a Superior Genetic Detection System

描述(申请人提供)：尽管所有生命系统的遗传信息的存储是从仅两个碱基对(A：T和G：C)的组织串中获得的，但额外的核酸碱基和碱基对在化学上是可能的。我们已经证明，编码第三个碱基对的另外两个碱基可以与自然界中的I四碱基、两碱基对系统一起应用于PCR。本项目将致力于利用这一新发现的六碱基聚合酶链式反应系统来改进和简化用于高度多重遗传分析的连接系统。三个碱基对的聚合酶链式反应技术将用于遗传分析系统，该系统首先使用连接来特定地连接序列特定的引物组，然后通过聚合酶链式反应来扩增这些连接的组。使用三个碱基对带来的额外复杂性将允许通过室温杂交来分析所建议的分析系统的产物。这种分析将实现比单独使用天然DNA的更高的复杂性和更好的辨别力。其目的是简化和改进六碱基聚合酶链式反应的遗传分析技术，以降低成本，提高稳健性和准确性。如果这个项目的第一阶段能够证明这一点，那么更多的化学应该受益于包括适体生产和DNA标记策略在内的六碱基聚合酶链式反应系统。

DESCRIPTION (provided by applicant): Although, the storage of genetic information for all living systems is derived from the organizational string of just two base pairs (A:T and G:C), additional nucleic acid bases and base-pairs are chemically possible. We have shown that two additional bases encoding a third base pair can be applied to PCR along with the I four-base, two-base-pair system of nature. This project will focus on using this newly discovered six-base PCR system to advance and simplify ligonucleotide ligation systems for highly multiplexed genetic analysis. The three-base-pair PCR technology will be used in a genetic analysis system which first uses ligation to specifically couple sequence-specific primer sets and then PCR to amplify those ligated sets. The additional complexity provided by using three base pairs will allow the products of the proposed analysis system to be analyzed by room temperature hybridization. This analysis will achieve higher complexity and better discrimination than that possible with natural DNA alone. The goal is to simplify and advance genetic analysis technology with six-base PCR in order to lower cost and increase robustness and accuracy. If this can be demonstrated in Phase I of this project, additional chemistries should benefit from a six-base PCR system including aptamer production and DNA tagging strategies.