SELECTION OF PRIMERS FOR POLYMERASE CHAIN-REACTION
SELECTION OF PRIMERS FOR POLYMERASE CHAIN-REACTION
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DOI：
10.1007/bf02789108
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发表时间：
1995-04-01
影响因子：
2.6
通讯作者：
RYCHLIK, W
中科院分区：
文献类型：
--
作者：
RYCHLIK, W
影响聚合酶链式反应质量的重要因素之一是引物的选择。一般来说，扩增产物越长，就越难选择有效的引物和设置合适的设计引物，而且一般来说，获得的DNA序列信息越多，找到最佳引物对的机会就越大。可以通过避免以下缺陷来设计有效的引物：引物-二聚体形成、自身互补性、引物的T-m太低和/或其内部稳定性曲线不正确。给出了亚克隆PCR产物、计算双链稳定性(预测二聚体形成强度)和设计简并引物的技巧。
One of the most important factors affecting tbe quality of PCR is the choice of primers. In general, the longer the PCR product the more difficult it is to select efficient primers and set appropriate designing primers, and in general, the more DNA sequence information is available, the better the chance of finding an optimal primer pair. Efficient primers can be designed by avoiding the following flaws: primer-dimer formation, self-complementarity, too low T-m of the primers, and/or their incorrect internal stability profile. Tips on subcloning PCR products, calculating duplex stability (predicting dimer formation strength), and designing degenerate primers are given.