A third base pair for the polymerase chain reaction: inserting isoC and isoG
A third base pair for the polymerase chain reaction: inserting isoC and isoG
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DOI：
10.1093/nar/gkh522
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发表时间：
2004-03-01
影响因子：
14.9
通讯作者：
Prudent, JR
中科院分区：
文献类型：
--
作者：
Johnson, SC;Sherrill, CB;Prudent, JR
另外两个碱基（异鸟苷和异胞嘧啶），产生第三个碱基对，已在PCR中实施。第三碱基对的酶保真度通过分子热力学熔融、化学裂解和分子信标来证明。当扩增不到15个含有多个非天然碱基对的靶标时，我们的结果证实了序列守恒。三个碱基对提供的额外序列空间允许构建分子工具，以实现比单独使用天然DNA更高的复杂性和更好的识别。
Two additional bases (isoguanosine and isocytosine), generating a third base pair, have been implemented in PCR. Enzyme fidelity for the third base pair is demonstrated using molecular thermodynamic melting, chemical cleavage and molecular beacons. When amplifying as few as 15 targets containing multiple non-natural base pairs with 40 cycles of amplification, our results confirm sequence conservation. The additional sequence space provided by three base pairs allows for the construction of molecular tools that achieve higher complexity and better discrimination than those possible with natural DNA alone.