Chandra Prafulla K, Wikel Stephen K
Center for Microbial Pathogenesis, MC3710, School of Medicine, University of Connecticut Health Center, Farmington, CT 06030-3710, USA.
Biol Proced Online. 2005;7:93-100. doi: 10.1251/bpo108. Epub 2005 Jun 17.
We have developed a simple and effective method (Lig-PCR) for monitoring ligation reactions using PCR and primers that are common to many cloning vectors. Ligation mixtures can directly be used as templates and the results can be analyzed by conventional gel electrophoresis. The PCR products are representative of the recombinant molecules created during ligation and the corresponding transformants. Orientation of inserts can also be determined using an internal primer. The usefulness of this method has been demonstrated using ligation mixtures of two cDNA's derived from the salivary glands of Aedes aegypti mosquitoes. The method described here is sensitive and easy to perform compared to currently available methods.
我们开发了一种简单有效的方法（连接聚合酶链反应，Lig-PCR），用于使用聚合酶链反应（PCR）和许多克隆载体通用的引物来监测连接反应。连接混合物可直接用作模板，结果可通过常规凝胶电泳进行分析。PCR产物代表连接过程中产生的重组分子及相应的转化体。插入片段的方向也可使用内部引物来确定。利用源自埃及伊蚊唾液腺的两个互补DNA（cDNA）的连接混合物，已证明了该方法的实用性。与目前可用的方法相比，这里描述的方法灵敏且易于操作。
