Borodina Tatiana A, Lehrach Hans, Soldatov Aleksey V
Max-Planck-Institut fur Molekulare Genetik, 14195 Berlin-Dahlem, Germany.
Anal Biochem. 2003 Jul 15;318(2):309-13. doi: 10.1016/s0003-2697(03)00250-1.
We describe here a method for the synthesis of oligonucleotides with block structure (padlock probes, primers for multiplex polymerase chain reaction (PCR), and ligation-independent cloning), based on the ligation of presynthesized parts by T4 DNA ligase. The advantages of this approach are: (i) suitability of the technology for any producer-from synthesis company to laboratory, (ii) high quality and adjustable scale of synthesis, and (iii) possibility of including any modified bases inexpensively in the common part of the oligonucleotide. Clear difference of sizes of products and substrates makes the synthesis amenable to automation. For large series of padlock probes, the price per one primer approaches the price of the locus-specific parts. We demonstrate the application of this method to two different tasks: preparative-scale production of padlock probes and small-scale synthesis of PCR primers.
我们在此描述一种合成具有块状结构的寡核苷酸的方法（锁式探针、多重聚合酶链式反应（PCR）引物以及不依赖连接的克隆），该方法基于通过T4 DNA连接酶连接预先合成的部分。这种方法的优点包括：（i）该技术适用于任何生产者——从合成公司到实验室；（ii）合成质量高且规模可调整；（iii）能够在寡核苷酸的通用部分廉价地包含任何修饰碱基。产物和底物在大小上的明显差异使得该合成适合自动化操作。对于大量的锁式探针，每一个引物的价格接近位点特异性部分的价格。我们展示了该方法在两项不同任务中的应用：锁式探针的制备规模生产以及PCR引物的小规模合成。
